Nematicidal aqueous suspension concentrate compositions

ABSTRACT

Provided herein are aqueous suspension concentrate compositions comprising biologically active 3,5-disubstituted-1,2,4-oxadiazoles or salts thereof that are useful, for example, in the control of nematodes. Nematodes are active, flexible, elongate organisms that live on moist surfaces or in liquid environments, including films of water within soil and moist tissues within other organisms. Many species of nematodes have evolved to be very successful parasites of plants and animals and, as a result, are responsible for significant economic losses in agriculture and livestock.

REFERENCE TO RELATED APPLICATIONS

This application is the United States National Stage Application ofInternational PCT Application Serial No. PCT/US2013/073128, filed Dec.4, 2013, and claims the benefit of U.S. Provisional Application Ser. No.61/733,239, filed Dec. 4, 2012, the entire contents of which areincorporated herein by reference.

FIELD

Provided herein are aqueous suspension concentrate compositionscomprising biologically active 3,5-disubstituted-1,2,4-oxadiazoles orsalts thereof that are useful, for example, in the control of nematodes.

BACKGROUND

Nematodes are active, flexible, elongate organisms that live on moistsurfaces or in liquid environments, including films of water within soiland moist tissues within other organisms. Many species of nematodes haveevolved to be very successful parasites of plants and animals and, as aresult, are responsible for significant economic losses in agricultureand livestock.

Plant parasitic nematodes can infest all parts of the plant, includingthe roots, developing flower buds, leaves, and stems. Plant parasitescan be classified on the basis of their feeding habits into a few broadcategories: migratory ectoparasites, migratory endoparasites, andsedentary endoparasites. Sedentary endoparasites, which include rootknot nematodes (Meloidogyne) and cyst nematodes (Globodera andHeterodera), can establish long-term infections within roots that may bevery damaging to crops.

There is an urgent need in the industry for effective, economical, andenvironmentally safe methods of controlling nematodes. Continuingpopulation growth, famines, and environmental degradation haveheightened concern for the sustainability of agriculture.

Recently, a class of 3,5-disubstituted-1,2,4-oxadiazoles has been shownto exhibit potent, broad spectrum nematicidal activity. See generallyU.S. Pat. No. 8,435,999 and U.S. Pat. No. 8,017,555, the contents ofwhich are expressly incorporated herein by reference. The3,5-disubstituted-1,2,4-oxadiazoles disclosed in U.S. Pat. No. 8,435,999and U.S. Pat. No. 8,017,555 are generally characterized by low watersolubility.

Two-phase suspension concentrates, which comprise solid particles of acompound suspended in an aqueous medium, are generally known in the art.In the context of seed treatment applications, suspension concentratesare known to offer several potential advantages, including high activeloading, ease of handling, and reduced toxicity and flammabilityassociated with solvents. The suspension concentrate compositions knownin the art, however, are also prone to instability and settling uponstorage, and may not provide a uniform distribution of the activenematicide compound in a manner that enhances bioavailability.

To be effective for use as a seed treatment composition, a nematicidalsuspension concentrate desirably satisfies several key requirements. Thenematicidal active ingredient must be effectively incorporated into asuspension having commercially acceptable storage stability. Thesuspension should exhibit acceptable storage stability over a widetemperature range and even where the nematicidal active ingredient ispresent in a high loading, which reduces the required volume of thecomposition and, therefore, reduces the expense of storage and shipping.The nematicidal active ingredient must also be amenable to transfer fromthe suspension concentrate to the surface of the seed, such that thedesired loading can be efficiently achieved. Moreover, followingapplication to the seed, it may be desirable for the nematicidal activeingredient to effectively migrate from the seed surface to the root zoneof the surrounding soil.

Accordingly, there remains a need in the art to develop compositionsthat enable the efficient use of the above-mentioned potent andeffective 3,5-disubstituted-1,2,4-oxadiazole nematicidal compounds inlarge-scale, commercial agricultural applications, particularly in seedtreatment applications, to protect against nematode infestations.

SUMMARY OF THE INVENTION

In one aspect, the present invention is therefore directed to anematicidal aqueous suspension concentrate composition, wherein thecomposition comprises a continuous aqueous phase comprising a dispersantcomponent, and a dispersed solid particulate phase comprising anematicidal component, the nematicidal component comprising a3,5-disubstituted-1,2,4-oxadiazole compound or a salt thereof, whereinthe median size of solid particulates in the dispersed solid particulatephase is less than about 10 μm.

In one embodiment, the present invention is directed to a nematicidalaqueous suspension concentrate composition as described above, whereinthe nematicidal component comprises a compound of Formula (I) or a saltthereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃.

In another embodiment, the present invention is directed to anematicidal aqueous suspension concentrate composition as describedabove, wherein the nematicidal component comprises a compound of Formula(II) or a salt thereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more with substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃.

Another aspect of the present invention is directed to methods ofpreparing the nematicidal aqueous suspension concentrate compositionsdescribed above. In one embodiment, the method comprises mixing thenematicidal compound, the dispersant, and water to form an aqueoussuspension; and wet milling the aqueous suspension to produce a milledsuspension having a reduced particle size.

Another aspect of the present invention is directed to methods ofprotecting the roots of a plant against damage by a nematode, the methodcomprising applying a nematicidal aqueous suspension concentratecomposition as described above the soil surrounding the root zone of aplant.

Another aspect of the present invention is directed to methods ofprotecting a seed and/or the roots of a plant grown from the seedagainst damage by a nematode, the method comprising treating a seed witha seed treatment composition, the seed treatment composition comprisinga nematicidal aqueous suspension concentrate composition as describedabove.

Another aspect of the present invention is directed to a seed that hasbeen treated with a seed treatment composition, the seed treatmentcomposition comprising a nematicidal aqueous suspension concentratecomposition as described above.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a representative photomicrograph of polymorphic Form I of3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIG. 2 depicts a representative photomicrograph of polymorphic Form IIof 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIG. 3 depicts a sample cyclic differential scanning calorimetry (DSC)thermogram from a cyclic DSC analysis conducted on3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole at a cooling rate of 30° C.per minute.

FIG. 4 depicts an X-ray diffraction (XRD) overlay of polymorphic Forms Iand II of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIGS. 5A and 5B depict XRD overlay results for polymorphic Forms I andII of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole, respectively.

FIG. 6 depicts the results of a powder XRD analysis of the Form Ipolymorph of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIG. 7 depicts the results of a powder XRD analysis of the Form IIpolymorph of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIG. 8 depicts a graphical XRD overlay of the competitive slurryexperiment between polymorphic Forms I and II of3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIGS. 9A through 9C depict the relevant DSC thermograms for polymorphicForm I, polymorphic Form II, and a mixture of polymorphic Forms I andII, respectively, of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole.

FIG. 10 depicts the results of an XRD analysis on samples of3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole material after 4 weeks ofstorage.

FIG. 11 depicts the XRD overlays of Forms I, II and the sample of FormII which showed signs of transformation to Form I.

DETAILED DESCRIPTION OF THE INVENTION

Provided herein are aqueous suspension concentrate nematicidalcompositions comprising 3,5-disubstituted-1,2,4-oxadiazoles and havingimproved effectiveness for seed treatment applications.

It has been discovered that the dispersibility of solid particulates ofthese generally hydrophobic, nematicidal compounds in an aqueous mediumcan be significantly increased through the application of millingtechniques that substantially reduce the mean and median particle sizecharacteristics of the dispersed solid phase, and by employing selecteddispersants. The reduced size of the solid particulates enables thepreparation of storage-stable, high-load suspension concentratecompositions. Increasing the aqueous dispersibility of these activenematicidal agents is highly beneficial, particularly in agriculturalapplications. For example, the compositions of the present invention maybe advantageously applied to seeds as a prophylactic treatment againstnematode infestation. Improved aqueous dispersibility provides for amore effective dispersion and more consistent loading of the nematicidalcompound during initial application of the composition to the seed. Inaddition, the improved aqueous dispersibility provided by the presentcompositions is beneficial during the post-planting stage, as it allowsthe nematicide to more effectively disperse throughout the hydrophilicenvironment in the soil surrounding the seed and, subsequently, the rootzone of the plant. Furthermore, it has been discovered that bycontrolling the particle size distribution of the nematicide particlesas described herein, the adhesion characteristics of the active compoundon the surface of the seeds allows for the efficient production oftreated seeds having the desired active loading, and later enhances thebioavailability of the active compound in the soil.

The aqueous suspension concentrate nematicidal compositions describedherein are sometimes referred to herein as “suspension concentratecompositions,” or more briefly as “suspension concentrates” or “thecomposition.” The suspension concentrate composition may also bereferred to herein as a “seed treatment composition,” particularly inthe context of seed treatment applications.

Nematicide

The aqueous compositions described herein generally comprise anematicide component comprising one or more3,5-disubstituted-1,2,4-oxadiazole compounds.

For example, in one embodiment, the nematicide component comprises acompound of Formula I or a salt thereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃.

In a more specific embodiment, the nematicide component comprises a3,5-disubstituted-1,2,4-oxadiazole of Formula Ia or a salt thereof,

wherein R₁ and R₅ are independently selected from the group consistingof hydrogen, CH₃, F, Cl, Br, CF₃ and OCF₃; R₂ and R₄ are independentlyselected from the group consisting of hydrogen, F, Cl, Br, and CF₃; R₃is selected from the group consisting of hydrogen, CH₃, CF₃, F, Cl, Br,OCF₃, OCH₃, CN, and C(H)O; R₇ and R₈ are independently selected fromhydrogen and F; R₉ is selected from the group consisting of hydrogen, F,Cl, CH₃, and OCF₃; and E is O, N or S. Typically, E is selected from thegroup consisting of O and S.

In another embodiment, the nematicide component comprises a compound ofFormula Ib or a salt thereof,

wherein R₁ and R₅ are independently selected from the group consistingof hydrogen, CH₃, F, Cl, Br, CF₃ and OCF₃; R₂ and R₄ are independentlyselected from the group consisting of hydrogen, F, Cl, Br, and CF₃; R₃is selected from the group consisting of hydrogen, CH₃, CF₃, F, Cl, Br,OCF₃, OCH₃, CN, and C(H)O; R₈ is selected from hydrogen and F; R₆ and R₉are independently selected from the group consisting of hydrogen, F, Cl,CH₃, and OCF₃; and E is N, O or S. Typically, E is selected from thegroup consisting of O and S.

In another embodiment, the nematicide component comprises a3,5-disubstituted-1,2,4-oxadiazole of Formula II or a salt thereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more with substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃.

In a more specific embodiment, the nematicide component comprises acompound of Formula IIa or a salt thereof,

wherein R₁ and R₅ are independently selected from the group consistingof hydrogen, CH₃, F, Cl, Br, CF₃ and OCF₃; R₂ and R₄ are independentlyselected from the group consisting of hydrogen, F, Cl, Br, and CF₃; R₃is selected from the group consisting of hydrogen, CH₃, CF₃, F, Cl, Br,OCF₃, OCH₃, CN, and C(H)O; R₇ and R₈ are independently selected fromhydrogen and F; R₉ is selected from the group consisting of hydrogen, F,Cl, CH₃, and OCF₃; and E is N, O or S. Typically, E is selected from thegroup consisting of O and S.

In another embodiment, the nematicide component comprises a compound ofFormula IIb or a salt thereof,

wherein R₁ and R₅ are independently selected from the group consistingof hydrogen, CH₃, F, Cl, Br, CF₃ and OCF₃; R₂ and R₄ are independentlyselected from the group consisting of hydrogen, F, Cl, Br, and CF₃; R₃is selected from the group consisting of hydrogen, CH₃, CF₃, F, Cl, Br,OCF₃, OCH₃, CN, and C(H)O; R₈ is selected from hydrogen and F; R₆ and R₉are independently selected from the group consisting of hydrogen, F, Cl,CH₃, and OCF₃; and E is N, O or S. Typically, E is selected from thegroup consisting of O and S.

In a preferred embodiment, the nematicidal component comprises a3,5-disubstituted-1,2,4-oxadiazole of Formula (Ia) or a salt thereof.Non-limiting examples of species include3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole of Formula (Ia-i),

3-(4-chlorophenyl)-5-(furan-2-yl)-1,2,4-oxadiazole of Formula (Ia-ii),

3-(4-chloro-2-methylphenyl)-5-(furan-2-yl)-1,2,4-oxadiazole of Formula(Ia-iii),

and 5-(furan-2-yl)-3-phenyl-1,2,4-oxadiazole of Formula (Ia-iv).

In another embodiment, the nematicidal component comprises a3,5-disubstituted-1,2,4-oxadiazole of Formula (Ib) or a salt thereofNon-limiting examples of species include3-(4-bromophenyl)-5-(furan-3-yl)-1,2,4-oxadiazole of Formula (Ib-i),

and 3-(2,4-difluorophenyl)-5-(thiophen-3-yl)-1,2,4-oxadiazole of Formula(Ib-ii).

In another embodiment, the nematicidal component comprises a3,5-disubstituted-1,2,4-oxadiazole of Formula (IIa) or a salt thereof.Non-limiting examples of species include3-(thiophen-2-yl)-5-(p-tolyl)-1,2,4-oxadiazole of Formula (IIa-i),

5-(3-chlorophenyl)-3-(thiophen-2-yl)-1,2,4-oxadiazole of Formula(IIa-ii),

and 5-(4-chloro-2-methylphenyl)-3-(furan-2-yl)-1,2,4-oxadiazole ofFormula (IIa-iii).

Polymorphs of the Nematicidal Compounds

The aqueous suspension concentrate composition can comprise any of thepolymorphic forms of the nematicidal compounds described herein.

Generally, polymorphism refers to the potential of a chemical entity toexist in different three-dimensional arrangements in the solid state.Different polymorphic forms of a compound can have different physicalproperties, including: solubility and dissolution rate; crystal shape;solid state stability; batch-to-batch manufacturing reproducibility;stability; ease of formulation; and bioavailability, among others. Indeciding which polymorph of a given compound is preferable for aspecific application, the relevant properties of each polymorph shouldbe determined and compared, so that the polymorph with the mostdesirable combination of attributes can be selected for use.

For example, it has been discovered that the nematicidal compound3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole, referred to herein as thecompound of Formula (Ia-i), exists in two distinct polymorphic forms,referred to herein as Form I and Form II. Form I is believed to be thethermodynamically stable form under ambient conditions, while Form II ismetastable at room temperature and pressure. The polymorphs areenantiotropically related. The transition temperature between the twoforms is believed to be approximately 102° C., wherein Form I is thestable form below the transition temperature, and Form II is the morethermodynamically stable form above that temperature.

Form I is believed to correspond to a dry crystalline polymorphic formof the compound. Generally, Form I does not appear to be prone tohydrate formation. Microscopic evaluation of Form I showed birefringentacicular to columnar shaped particles ranging from approximately 50 to100 microns in length. FIG. 1 shows the representative photomicrographat room temperature.

Form II is also believed to correspond to a dry crystalline polymorphicform of the compound. Microscopic evaluation of Form II showedbirefringent acicular, columnar, and flake shaped particles ranging fromapproximately 25 to 150 microns in length. FIG. 2 shows therepresentative photomicrograph at room temperature.

Generally, the aqueous suspension concentrate composition can compriseany of the polymorphic forms of the nematicidal compounds describedherein. For example, in one embodiment, the suspension concentratecomposition comprises polymorphic Form I of the compound of Formula(Ia-i). In another embodiment, the suspension concentrate compositioncomprises polymorphic Form II of the compound of Formula (Ia-i).Mixtures of more than one polymorph are also considered to be within thescope of the invention. For example, in one embodiment, the suspensionconcentrate composition comprises a mixture of polymorphic forms I andII of the compound of Formula (Ia-i).

Concentration

The suspension concentrate composition in some embodiments comprises atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 35%, at least about 40%, atleast about 45%, or at least about 50% by weight of the nematicidecomponent comprising one or more active nematicidal compounds asdescribed above. In one embodiment, the suspension concentratecomposition comprises at least about 40% by weight of the nematicidecomponent. In some embodiments, the suspension concentrate compositioncomprises at least about 45% by weight of the nematicide component, oreven higher (e.g., at least about 50% by weight).

The suspension concentrate composition comprises the nematicidecomponent in a concentration of at least about 100 g/L, at least about200 g/L, at least about 250 g/L, at least about 300 g/L, at least about350 g/L, at least about 400 g/L, at least about 450 g/L, at least about500 g/L, at least about 550 g/L, at least about 600 g/L, at least about650 g/L, or at least about 700 g/L. The nematicide concentration rangesfrom about 400 g/L to about 700 g/L, from about 450 g/L to about 750g/L, or from about 450 g/L to about 700 g/L.

Particle Size

The suspension concentrate compositions of the present inventioncomprise a continuous aqueous phase and a dispersed solid phasecomprising solid particulates of the nematicide component as describedherein. The solid nematicidal particulates have a particle sizedistribution selected to enhance dispersibility of the particlessuspended in the composition and improve the stability of the suspensionconcentrate composition.

It has been discovered, however, that further reductions in particlesize provide a number of benefits, including improved adhesioncharacteristics of the 3,5-disubstituted-1,2,4-oxadiazole compounds whenthe composition is applied as a seed treatment. The particle sizereduction described herein provides enhanced adhesion of the nematicidalactive ingredient to the seed surface when the composition is applied asa seed treatment and thereby allows for efficient production of treatedseeds having a uniform active loading. Furthermore, and without beingbound to a particular theory, it is believed that further reducing theparticulate size of the 3,5-disubstituted-1,2,4-oxadiazole compoundsfacilitates improved dispersibility of the solid nematicidal activewithin the aqueous environment of the root zone after planting thetreated seed in the soil. Dispersion of the nematicide throughout thesurrounding root zone helps prevent soil nematodes from coming intocontact with the seed and, later, the newly formed roots of the plantemerging from the seed, and ultimately manifests as an improvement innematicidal efficacy (i.e., a reduction in plant damage attributable tonematodes).

In the preparation of suspension concentrates, there are considerableenergy costs and time requirements associated with reducing the particlesize of the solid phase. These costs tend to increase significantly asthe particle size decreases. Accordingly, efficient production ofsuspension concentrates must take into account the additional costs andbenefits associated with the particle size reduction step.

Accordingly, the particle size characteristics of the dispersed solidphase of the suspension concentrate composition comprising the3,5-disubstituted-1,2,4-oxadiazole compounds described above areselected so as to not only provide a stable suspension, but also toallow for efficient production of treated seeds having a uniform activeloading and enhanced nematicidal efficacy. More particularly, thedispersed solid phase of the suspension concentrate has a medianparticle size less than about 50 μm, less than 30 μm, less than 20 μm,less than 10 μm, less than about 5 μm, less than about 4 μm, less thanabout 3 μm, less than about 2 μm, or less than about 1 μm. Thesuspension concentrate composition typically has a median particle sizefalling within the range of from about 0.5 μm to about 10 μm, from about1 μm to about 5 μm, from about 1 μm to about 4 μm, from about 1 μm toabout 3 μm, or from about 1 μm to about 2 μm. In some embodiments, themedian particle size falls within the range of from about 0.5 μm toabout 5 μm, from about 0.5 μm to about 4 μm, from about 0.5 μm to about3 μm, from about 0.5 μm to about 2 μm, or from about 0.5 μm to about 1μm. In one embodiment, the median particle size falls within the rangeof from about 1 μm to about 2 μm.

The dispersed solid phase of the suspension concentrate compositiontypically has a mean particle size less than about 20 μm, less thanabout 10 μm, less than about 5 μm, less than about 4 μm, less than about3 μm, less than about 2 μm, or less than about 1 μm. The mean particlesize typically falls within the range of from about 0.5 μm to about 20μm, from about 0.5 μm to about 10 μm, from about 1 μm to about 5 μm,from about 1 μm to about 4 μm, from about 1 μm to about 3 μm, or fromabout 1 μm to about 2 μm. In some embodiments, the mean particle sizefalls within the range of from about 0.5 μm to about 5 μm, from about0.5 μm to about 4 μm, from about 0.5 μm to about 3 μm, from about 0.5 μmto about 2 μm, or from about 0.5 μm to about 1 μm.

The mean and/or median particle size of the solid particulates in thedispersed phase can be determined by means known in the art, includinglaser diffraction particle size analysis. A non-limiting example of asuitable apparatus for determining the particle size characteristics ofthe solid particulates is a BECKMAN COULTER LS Particle Size Analyzer(model LS 13 320).

The dispersed solid phase of the suspension concentrate typically has apolydispersity index, defined as the arithmetic mean particle sizedivided by the median particle size, of less than about 10. In someembodiments, the polydispersity index is less than about 5, less thanabout 2, or less than about 1.5. The polydispersity index typicallyfalls within the range of from about 1 to about 2.

Dispersant

The suspension concentrate composition additionally comprises adispersant component comprising one or more dispersants selected toenhance dispersibility of the solid particles suspended in thecomposition and improve the stability of the suspension concentratecomposition. The dispersant may be selected from non-ionic dispersants,anionic dispersants, or cationic dispersants.

In a preferred embodiment, the dispersant is anionic. Examples ofanionic dispersants include alkyl sulfates, alcohol sulfates, alcoholether sulfates, alpha olefin sulfonates, alkylaryl ether sulfates,arylsulfonates, alkylsulfonates, alkylaryl sulfonates, sulfosuccinates,mono- or diphosphate esters of polyalkoxylated alkyl alcohols or alkylphenols, mono- or disulfosuccinate esters of alcohols or polyalkoxylatedalkanols, alcohol ether carboxylates, phenol ether carboxylates.

In one embodiment, the dispersant is an alkylaryl sulfonate. Alkylarylsulfonates have been found to be effective at forming a stable aqueoussuspension comprising the 3,5-disubstituted-1,2,4-oxadiazole compoundsused in the practice of the present invention, particularly at highconcentrations of the nematicidal active ingredient.

Non-limiting examples of commercially available anionic dispersantsinclude sodium dodecylsulfate (Na-DS, SDS), MORWET D-425 (a sodium saltof alkyl naphthalene sulfonate condensate, available from Akzo Nobel),MORWET D-500 (a sodium salt of alkyl naphthalene sulfonate condensatewith a block copolymer, available from Akzo Nobel), sodiumdodecylbenzene sulfonic acid (Na-DBSA) (available from Aldrich),diphenyloxide disulfonate, naphthalene formaldehyde condensate, DOWFAX(available from Dow), dihexylsulfosuccinate, and dioctylsulfosuccinate.For example, the anionic dispersant may comprise an alkyl naphthalenesulfonate condensate or a salt thereof.

Examples of non-ionic dispersants include sorbitan esters, ethoxylatedsorbitan esters, alkoxylated alkylphenols, alkoxylated alcohols, blockcopolymer ethers, and lanolin derivatives. In accordance with oneembodiment, the dispersant comprises an alkylether block copolymer

Non-limiting examples of commercially available non-ionic dispersantsinclude SPAN 20, SPAN 40, SPAN 80, SPAN 65, and SPAN 85 (available fromAldrich); TWEEN 20, TWEEN 40, TWEEN 60, TWEEN 80, and TWEEN 85(available from Aldrich); IGEPAL CA-210, IGEPAL CA-520, IGEPAL CA-720,IGEPAL CO-210, IGEPAL CO-520, IGEPAL CO-630, IGEPAL CO-720, IGEPALCO-890, and IGEPAL DM-970 (available from Aldrich); Triton X-100(available from Aldrich); BRIJ S10, BRIJ S20, BRIJ 30, BRIJ 52, BRIJ 56,BRIJ 58, BRIJ 72, BRIJ 76, BRIJ 78, BRIJ 92V, BRIJ 97, and BRIJ 98(available from Aldrich); PLURONIC L-31, PLURONIC L-35, PLURONIC L-61,PLURONIC L-81, PLURONIC L-64, PLURONIC L-121, PLURONIC 10R5, PLURONIC17R4, and PLURONIC 31R1 (available from Aldrich); Atlas G-5000 and AtlasG-5002 L (available from Croda); ATLOX 4912 and ATLOX 4912-SF (availablefrom Croda); and SOLUPLUS (available from BASF), LANEXOL AWS (availablefrom Croda).

Non-limiting examples of cationic dispersants include mono alkylquaternary amine, fatty acid amide surfactants, amidoamine, imidazoline,and polymeric cationic surfactants.

The suspension concentrate composition comprises from about 0.5% about20%, from about 0.5% to about 10%, from about 0.5% to about 5%, or fromabout 0.5% to about 8% of the dispersant component by weight. In oneembodiment, the composition comprises the dispersant in an amount offrom about 0.5% to about 5% by weight.

The suspension concentrate composition may comprise the dispersant in aconcentration of at least about 5 g/L, at least about 10 g/L, at leastabout 15 g/L, at least about 20 g/L, at least about 25 g/L, at leastabout 30 g/L, at least about 35 g/L, at least about 40 g/L, at leastabout 45 g/L, or at least about 50 g/L. In some embodiments, thedispersant is present in a concentration of from about 1 to about 100g/L, from about 5 to about 75 g/L, or more typically from about 20 toabout 50 g/L.

In some embodiments, the suspension concentrate composition comprises adispersant component comprising a primary dispersant in combination withone or more secondary dispersants. The secondary dispersant may also bereferred to herein as a wetting agent. In one embodiment, the secondarydispersant comprises an alkylether block copolymer.

In one embodiment, the secondary dispersant is non-ionic when used inconjunction with an ionic primary dispersant. For example, in someembodiments, the dispersant component comprises a mixture of an anionicprimary dispersant (described above) and a non-ionic (described above)secondary dispersant. In other embodiments, the dispersant componentcomprises a mixture of a cationic primary dispersant and a non-ionicsecondary dispersant. In accordance with another embodiment, it has beenfound that the pairing of an anionic primary dispersant with a non-ionicsecondary dispersant, in particular, imparts improved stability to theaqueous suspension concentrates described herein.

The secondary dispersant typically comprises from about 0.05% to about10%, from about 0.5% to about 5%, from about 1% to about 5%, from about1% to about 4%, or from about 1% to about 2.5% by weight of thecomposition.

The composition typically comprises a ratio of primary dispersant tosecondary dispersant, on a weight basis, of from about 1:1 to about10:1, from about 1:1 to about 5:1, and from about 2:1 to about 3:1.

Dendrimers

In some embodiments, the composition may further comprise one or morefunctionalized dendrimers to enhance the efficacy and/or stability ofthe composition. Non-limiting examples of classes of functionalizeddendrimers include poly(amidoamine) (PAMAM, Generations 0-7),poly(amidoamine-organosilicone) (PAMAMOS), poly(propylene imidine) (PPI,Generations 0-5), poly(benzylethers) (Frechet-type), Arobols (Newkometype), poly(phenylacetylenes) and surface engineered dendrimers (e.g.PEGylated dendrimers, glycodendrimers, peptide funtionalized dendrimers,and galabiose-functionalized dendrimers). In some embodiments, thedendrimers comprise at least about 0.1% and up to 10% or more, or fromabout 1% to about 10% by weight of the composition.

Antifreeze Agents

In some embodiments, the composition may further comprise one or moreantifreeze agents. In one embodiment, the antifreeze agent is analcohol. Non-limiting examples of antifreeze agents include ethyleneglycol, propylene glycol, butanediol, pentanediol, mannitol, sorbitol,and glycerol (glycerin).

The suspension concentrate composition may comprise the antifreeze agentin a concentration of at least about 5 g/L, at least about 10 g/L, atleast about 15 g/L, at least about 20 g/L, at least about 30 g/L, atleast about 40 g/L, at least about 50 g/L, at least about 60 g/L, atleast about 70 g/L, or at least about 80 g/L. The antifreeze agent istypically present in a concentration of from about 1 to about 150 g/L,from about 10 to about 100 g/L, or more typically from about 20 to about80 g/L.

Antifoam Agents

In some embodiments, the composition may further comprise one or moreantifoam agents. Examples of antifoam agents include organosilicone orsilicone-free compounds. Non-limiting examples of commercially availableantifoam products include Break-Thru OE441 (available from Evonik),Break-Thru AF9905 (available from Evonik), AGNIQUE DF 6889 (availablefrom Cognis), AGNIQUE DFM 111S (available from Cognis), BYK-016(available from BYK), FG-10 antifoam emulsion (available from DowCorning), 1520-US (available from Dow Corning), 1510-US (available fromDow Corning), SAG 1538 (available from Momentive), and SAG 1572(available from Momentive).

Buffer

In some embodiments, the composition may comprise a buffer solution thathelps maintain the pH within a desired range. It has been discoveredthat, at a pH greater than about 10, wet milling and/or ball milling thenematicidal component described herein results in excessive clumpingand/or agglomeration making the particle size reduction difficult,instability or degradation of the nematicidal component, and/orinstability or degradation of the other suspension concentratecomponents described herein. As a result, a pH buffer may be selected toprovide an aqueous suspension concentrate composition having a pH ofless than 10, from about 5 to about 9, from about 6 to about 8, or about7. Buffer solutions suitable for a variety of pH ranges are generallyknown in the art.

Thickener

In some embodiments, the composition may comprise a thickener (referredto hereinafter as “stabilizer”) component. Examples of stabilizersinclude anionic polysaccharides and cellulose derivatives. In someembodiments, the stabilizer comprises a clay or a silica, or a colloidalhydrophilic silica. Non-limiting examples of commercially availablestabilizers include KELZAN CC (available from Kelco), methyl cellulose,carboxymethylcellulose and 2-hydroxyethylcellulose,hydroxymethylcellulose, kaolin, and microcrystalline cellulose. Anon-limiting example of a commercially available colloidal hydrophilicsilica is AEROSIL (available from Evonik).

The stabilizer component typically comprises from about 0.05% to about10% by weight of the composition. For example, in some embodiments, thestabilizer component comprises from about 0.1% to about 5%, from about0.1% to about 2%, or from about 0.1% to about 1% by weight of thecomposition.

Crystal Growth Inhibitor

In some embodiments, the composition may comprise a crystal growthinhibitor. Examples of crystal growth inhibitors include acryliccopolymers, polyethylene glycol, polyethylene glycol hydrogenated castoroil and combinations. Non-limiting examples of commercially availablecrystal growth inhibitor include ATLOX 4913 (available from Croda).

The crystal growth inhibitor component may comprise from about 1% toabout 10% by weight of the composition.

Co-Solvent

In some embodiments, the composition may further comprise a co-solventin addition to water. Non-limiting examples of co-solvents that can beused include, ethyl lactate, methyl soyate/ethyl lactate co-solventblends (e.g., STEPOSOL, available from Stepan), isopropanol, acetone,1,2-propanediol, n-alkylpyrrolidones (e.g., the AGSOLEX series,available from ISP), a petroleum based-oil (e.g., AROMATIC series andSOLVESSO series available from Exxon Mobil), isoparaffinic fluids (e.g.ISOPAR series, available from Exxon Mobil), cycloparaffinic fluids (e.g.NAPPAR 6, available from Exxon Mobil), mineral spirits (e.g. VARSOLseries available from Exxon Mobil), and mineral oils (e.g., paraffinoil).

Non-limiting examples of preferred commercially available organicsolvents include pentadecane, ISOPAR M, and ISOPAR V and ISOPAR L(available from Exxon Mobil).

Rheology Modifying Agent

In some embodiments, the composition may further comprise one or morerheology modifying agents.

Examples of rheology modifying agents include humic acid salts, fulvicacid salts, humin, and lignin salts.

In one embodiment, the rheology modifying agent is the sodium orpotassium salt of humic acid. Generally, a humic substance is oneproduced by biodegradation of dead organic matter, particularly deadplant matter (e.g., lignin). With respect to the compositions of thepresent invention, it has been discovered that compositions comprising ahumic acid exhibit a lower viscosity than similarly-loaded compositionsin the absence of a humic acid. Fulvic acids, which are humic acids oflower molecular weight and higher oxygen content than other humic acids,are used in some embodiments.

Additional Excipients

In some embodiments, composition comprises one or more additionalexcipients that improve the adhesion of the composition to the seed,provide a visual indication of successful coating (e.g., coloringagents), or otherwise impart improved characteristics to the coating.

Biocidal Agents

In some embodiments, the composition may further comprise one or morebiocidal agents. Typically, a biocidal component is included to preventfungal and/or bacterial growth within the suspension concentratecomposition, particularly when the composition is placed into storage.Examples of biocidal agents include dichlorophen or benzyl alcoholhemiformal based compounds, benzoisothiazolinones and rhamnolipids.Non-limiting examples of commercially available biocidal agents includeACTICIDE (available from THOR), PROXEL (available from Arch Chemical),and ZONIX (available from Jeneil).

Additional Active Ingredients

In some embodiments, the composition may be formulated, mixed in a seedtreater tank or combined on the seed by overcoating with one or moreadditional active ingredients in combination with the nematicidal3,5-disubstituted-1,2,4-oxadiazoles described herein.

The additional active ingredient may be, for example, an additionalpesticide. The pesticide may be, for example, an insecticide, afungicide, an herbicide, or an additional nematicide.

Non-limiting examples of insecticides and nematicides includecarbamates, diamides, macrocyclic lactones, neonicotinoids,organophosphates, phenylpyrazoles, pyrethrins, spinosyns, syntheticpyrethroids, tetronic and tetramic acids. In particular embodimentsinsecticides and nematicides include abamectin, aldicarb, aldoxycarb,bifenthrin, carbofuran, chlorantraniliprole, clothianidin, clothianidinand a Bacillus firmus, cyantraniliprole, cyfluthrin, cyhalothrin,cypermethrin, deltamethrin, dinotefuran, emamectin, ethiprole,fenamiphos, fipronil, flubendiamide, fluopyram, fosthiazate,imidacloprid, ivermectin, lambda-cyhalothrin, milbemectin, nitenpyram,oxamyl, permethrin, spinetoram, spinosad, spirodichlofen, spirotetramat,tefluthrin, thiacloprid, thiamethoxam, and thiodicarb,

Non-limiting examples of useful fungicides include aromatichydrocarbons, benzimidazoles, benzthiadiazole, carboxamides, carboxylicacid amides, morpholines, phenylamides, phosphonates, quinone outsideinhibitors (e.g. strobilurins), thiazolidines, thiophanates, thiophenecarboxamides, and triazoles. Particular examples of fungicides includeacibenzolar-S-methyl, azoxystrobin, benalaxyl, bixafen, boscalid,carbendazim, cyproconazole, dimethomorph, epoxiconazole, fluopyram,fluoxastrobin, flutianil, flutolanil, fluxapyroxad, fosetyl-A1,ipconazole, isopyrazam, kresoxim-methyl, mefenoxam, metalaxyl,metconazole, myclobutanil, orysastrobin, penflufen, penthiopyrad,picoxystrobin, propiconazole, prothioconazole, pyraclostrobin, sedaxane,silthiofam, tebuconazole, thifluzamide, thiophanate, tolclofos-methyl,trifloxystrobin, and triticonazole.

Non-limiting examples of herbicides include ACCase inhibitors,acetanilides, AHAS inhibitors, carotenoid biosynthesis inhibitors, EPSPSinhibitors, glutamine synthetase inhibitors, PPO inhibitors, PS IIinhibitors, and synthetic auxins, Particular examples of herbicidesinclude acetochlor, clethodim, dicamba, flumioxazin, fomesafen,glyphosate, glufosinate, mesotrione, quizalofop, saflufenacil,sulcotrione, and 2,4-D.

Additional actives may also comprise substances such as, biologicalcontrol agents, microbial extracts, natural products, plant growthactivators or plant defense agents. Non-limiting examples of biologicalcontrol agents include bacteria, fungi, beneficial nematodes, andviruses.

In certain embodiments, the biological control agent can be a bacteriumof the genus Actinomycetes, Agrobacterium, Arthrobacter, Alcaligenes,Aureobacterium, Azobacter, Beijerinckia, Brevibacillus, Burkholderia,Chromobacterium, Clostridium, Clavibacter, Comomonas, Corynebacterium,Curtobacterium, Enterobacter, Flavobacterium, Gluconobacter,Hydrogenophage, Klebsiella, Methylobacterium, Paenibacillus, Pasteuria,Phingobacterium, Photorhabdus, Phyllobacterium, Pseudomonas, Rhizobium,Serratia, Stenotrophomonas, Variovorax, and Xenorhadbus. In particularembodiments the bacteria is selected from the group consisting ofBacillus amyloliquefaciens, Bacillus cereus, Bacillus firmus, Bacillus,lichenformis, Bacillus pumilus, Bacillus sphaericus, Bacillus subtilis,Bacillus thuringiensis, Chromobacterium suttsuga, Pasteuria penetrans,Pasteuria usage, and Pseudomona fluorescens.

In certain embodiments the biological control agent can be a fungus ofthe genus Alternaria, Ampelomyces, Aspergillus, Aureobasidium,Beauveria, Colletotrichum, Coniothyrium, Gliocladium, Metarhisium,Muscodor, Paecilonyces, Trichoderma, Typhula, Ulocladium, andVerticilium. In particular embodiments the fungus is Beauveria bassiana,Coniothyrium minitans, Gliocladium virens, Muscodor albus, Paecilomyceslilacinus, or Trichoderma polysporum.

In further embodiments the biological control agents can be plant growthactivators or plant defense agents including, but not limited to harpin,Reynoutria sachalinensis, jasmonate, lipochitooligosaccharides,gibberellic acid, and isoflavones.

Methods of Preparation

Another aspect of the present invention is directed to methods ofpreparing the nematicidal suspension concentrate compositions describedherein.

As described above, it has been discovered that significant benefits inthe aqueous dispersibility of 3,5-disubstituted-1,2,4-oxadiazoles can beobtained and other advantages realized by reducing the particulate sizeof the solid phase in the suspension concentrate composition. Generally,the particulate size of the nematicide component may be reduced by anymethod known in the art. In accordance with one preferred embodiment,the particle size of the nematicide component is reduced by wet milling.Additionally, air milling, high pressure homogenization, spinning disc,grinding and solvent evaporation techniques can be used to reduce theparticle size of the nematicide component.

Typically, the first step in the process comprises a pre-milling stepwherein the nematicidal component comprising one or more activenematicidal compounds is combined with water and agitated to form anaqueous suspension. Typically, the dispersant is also added to theaqueous suspension prior to the particle size reduction step and acts asa wet-milling aid. Other optional components which may be added to theaqueous suspension before the particle size reduction step include asecondary dispersant and/or an antifreeze agent, each of which may beselected as described above. Additionally, in one embodiment, a buffersolution is added to the suspension prior to the particle size reductionstep; as discussed above, the pH of the suspension during the particlesize reduction step is preferably less than 10 in order to minimizeexcessive clumping and/or agglomeration making the particle sizereduction difficult, instability or degradation of the nematicidalcomponent, and/or instability or degradation of the other suspensionconcentrate components described herein.

The aqueous suspension is then wet-milled to obtain a suspensionconcentrate having the desired particle size distribution as describedabove. The wet-milling process may be carried out using techniques andapparatus known in the art. Ball milling is a particularly preferredtechnique, wherein the aqueous suspension is placed inside a rotatingcylinder containing grinding media. The grinding media are preferablyselected from the group consisting of stainless steel beads, zirconiumoxide beads, glass beads and ceramic beads. Non-limiting examples ofsuitable ball milling apparatus include a SIZEGVARI ATTRITOR millingsystem made by UNION PROCESS, and a MINI ZETA II milling machine made byNetzsch.

The wet-milling step typically produces a fine suspension comprising adispersed solid phase having a particle size distribution characterizedby the median and mean particle sizes and polydispersity index describedabove. Using laser diffraction particle size analysis or other suitablemeans, the duration and intensity of the wet-milling operation iscontrolled to provide a suspension concentrate composition having thedesired particle size characteristics.

Following the particle size reduction, the milled aqueous suspension maybe combined with an optional stabilizer component and/or one or moreadditional biocidal agents, each of which may be selected as describedabove.

Storage Stability

In one embodiment, the aqueous suspension concentrate compositiondescribed herein exhibits commercially acceptable storage stabilityacross a wide range of temperatures and environmental conditions. Inthis context, storage stability is generally defined as the absence ofsedimentation and the lack of any significant change in the rheologicalproperties of the composition (e.g., viscosity). Commercially acceptablestorage stability can be reliably achieved by selecting the variouscomponents of the aqueous suspension concentrate, particularly theprimary dispersant, optional secondary dispersant, and/or optionalstabilizer component, in accordance with the respective embodimentsdescribed in detail above. The suspension concentrate composition may bestorage-stable at 25° C. for at least about 1 week, at least about 2weeks, at least about 1 month, at least about 2 months, at least about 3months, at least about 6 months, at least about 12 months or at leastabout 18 months.

Methods of Application

Another aspect of the present invention is directed to methods forprotecting the roots of a plant against damage by nematodes.

Application to Seeds

In one embodiment, the method comprises protecting a seed, and/or theroots of a plant grown from the seed, against damage by a nematode bytreating the seed with a seed treatment composition described herein anddiluted as necessary to attain the desired nematicide compound loadingon the treated seeds.

The methods described herein can be used in connection with any speciesof plant and/or the seeds thereof. In preferred embodiments, however,the methods are used in connection with seeds of plant species that areagronomically important. In particular, the seeds can be of corn,peanut, canola/rapeseed, soybean, cucurbits, crucifers, cotton, beets,rice, sorghum, sugar beet, wheat, barley, rye, sunflower, tomato,sugarcane, tobacco, oats, as well as other vegetable and leaf crops. Insome embodiments, the seed is corn, soybean, or cotton seed. The seedmay be a transgenic seed from which a transgenic plant can grow andincorporate a transgenic event that confers, for example, tolerance to aparticular herbicide or combination of herbicides, increased diseaseresistance, enhanced tolerance to stress and/or enhanced yield.Transgenic seeds include, but are not limited to, seeds of corn, soybeanand cotton.

In one embodiment, the treatment composition is applied to the seedprior to sowing the seed so that the sowing operation is simplified. Inthis manner, seeds can be treated, for example, at a central locationand then dispersed for planting. This permits the person who plants theseeds to avoid the complexity and effort associated with handling andapplying the seed treatment compositions, and to merely handle and plantthe treated seeds in a manner that is conventional for regular untreatedseeds.

The seed treatment composition can be applied to seeds by any standardseed treatment methodology, including but not limited to mixing in acontainer (e.g., a bottle or bag), mechanical application, tumbling,spraying, immersion, and solid matrix priming Seed coating methods andapparatus for their application are disclosed in, for example, U.S. Pat.Nos. 5,918,413, 5,891,246, 5,554,445, 5,389,399, 5,107,787, 5,080,925,4,759,945 and 4,465,017, among others. Any conventional active or inertmaterial can be used for contacting seeds with the seed treatmentcomposition, such as conventional film-coating materials including butnot limited to water-based film coating materials.

For example, in one embodiment, a seed treatment composition can beintroduced onto or into a seed by use of solid matrix priming. Forexample, a quantity of the seed treatment composition can be mixed witha solid matrix material and then the seed can be placed into contactwith the solid matrix material for a period to allow the seed treatmentcomposition to be introduced to the seed. The seed can then optionallybe separated from the solid matrix material and stored or used, or themixture of solid matrix material plus seed can be stored or planteddirectly. Solid matrix materials which are useful in the presentinvention include polyacrylamide, starch, clay, silica, alumina, talc,mica, soil, sand, polyurea, polyacrylate, or any other material capableof absorbing or adsorbing the seed treatment composition for a time andreleasing the nematicide of the seed treatment composition into or ontothe seed. It is useful to make sure that the nematicide and the solidmatrix material are compatible with each other. For example, the solidmatrix material should be chosen so that it can release the nematicideat a reasonable rate, for example over a period of minutes, hours, days,or weeks.

Imbibition is another method of treating seed with the seed treatmentcomposition. For example, a plant seed can be directly immersed for aperiod of time in the seed treatment composition. During the period thatthe seed is immersed, the seed takes up, or imbibes, a portion of theseed treatment composition. Optionally, the mixture of plant seed andthe seed treatment composition can be agitated, for example by shaking,rolling, tumbling, or other means. After imbibition, the seed can beseparated from the seed treatment composition and optionally dried, forexample by patting or air drying.

The seed treatment composition may be applied to the seeds usingconventional coating techniques and machines, such as fluidized bedtechniques, the roller mill method, rotostatic seed treaters, and drumcoaters. Other methods, such as spouted beds may also be useful. Theseeds may be pre-sized before coating. After coating, the seeds aretypically dried and then transferred to a sizing machine for sizing.Such procedures are generally known in the art.

If the seed treatment composition is applied to the seed in the form ofa coating, the seeds can be coated using a variety of methods known inthe art. For example, the coating process can comprise spraying the seedtreatment composition onto the seed while agitating the seed in anappropriate piece of equipment such as a tumbler or a pan granulator.

In one embodiment, when coating seed on a large scale (for example acommercial scale), the seed coating may be applied using a continuousprocess. Typically, seed is introduced into the treatment equipment(such as a tumbler, a mixer, or a pan granulator) either by weight or byflow rate. The amount of treatment composition that is introduced intothe treatment equipment can vary depending on the seed weight to becoated, surface area of the seed, the concentration of the nematicideand/or other active ingredients in the treatment composition, thedesired concentration on the finished seed, and the like. The treatmentcomposition can be applied to the seed by a variety of means, forexample by a spray nozzle or revolving disc. The amount of liquid istypically determined by the assay of the formulation and the requiredrate of active ingredient necessary for efficacy. As the seed falls intothe treatment equipment the seed can be treated (for example by mistingor spraying with the seed treatment composition) and passed through thetreater under continual movement/tumbling where it can be coated evenlyand dried before storage or use.

In another embodiment, the seed coating may be applied using a batchprocess. For example, a known weight of seeds can be introduced into thetreatment equipment (such as a tumbler, a mixer, or a pan granulator). Aknown volume of seed treatment composition can be introduced into thetreatment equipment at a rate that allows the seed treatment compositionto be applied evenly over the seeds. During the application, the seedcan be mixed, for example by spinning or tumbling. The seed canoptionally be dried or partially dried during the tumbling operation.After complete coating, the treated sample can be removed to an area forfurther drying or additional processing, use, or storage.

In an alternative embodiment, the seed coating may be applied using asemi-batch process that incorporates features from each of the batchprocess and continuous process embodiments set forth above.

In still another embodiment, seeds can be coated in laboratory sizecommercial treatment equipment such as a tumbler, a mixer, or a pangranulator by introducing a known weight of seeds in the treater, addingthe desired amount of seed treatment composition, tumbling or spinningthe seed and placing it on a tray to thoroughly dry.

In another embodiment, seeds can also be coated by placing the knownamount of seed into a narrow neck bottle or receptacle with a lid. Whiletumbling, the desired amount of seed treatment composition can be addedto the receptacle. The seed is tumbled until it is coated with thetreatment composition. After coating, the seed can optionally be dried,for example on a tray.

In some embodiments, the treated seeds may also be enveloped with a filmovercoating to protect the nematicidal coating. Such overcoatings areknown in the art and may be applied using conventional fluidized bed anddrum film coating techniques. The overcoatings may be applied to seedsthat have been treated with any of the seed treatment techniquesdescribed above, including but not limited to solid matrix priming,imbibition, coating, and spraying, or by any other seed treatmenttechnique known in the art.

Application to Soil

In another aspect of the present invention, the nematicidal treatmentcomposition, diluted as necessary to attain the desired nematicidecompound loading, is directly applied to the soil surrounding the rootzone of a plant. The application may be performed using any method orapparatus known in the art, including pressurized spray application tothe soil surface or injected in the planting furrow, as well aschemigation via overhead sprinkler or drip systems, transplant watertreatments, and plant or root dips prior to planting. The rates used forthe suspension concentrate formulations for soil application may require0.5 to 2 kgs per hectare on a broadcast basis (rate per treated area ifbroadcast or banded).

Treated Seeds

Another aspect of the present invention is directed to a seed that hasbeen treated with a nematicidal seed treatment composition as describedherein. Typically, the seed has been treated with the seed treatmentcomposition using one of the seed treatment methods set forth above,including but not limited to solid matrix priming, imbibition, coating,and spraying. The seed may be of any plant species, as described above.

Typically, the treated seeds comprise the nematicidal compound in anamount of at least about 0.05 mg/seed, more typically from about 0.05 toabout 1 mg/seed, and even more typically from about 0.05 to about 0.5mg/seed.

In some embodiments, wherein the composition comprises a paraffinichydrocarbon solvent, the loading of active ingredient per treated seedcan be significantly reduced without compromising nematicidal efficacy.For example, when the seed treatment composition comprises a paraffinichydrocarbon solvent, the treated seeds may comprise the nematicidalcompound in an amount of less than about 0.2 mg/seed, in an amount ofabout 0.1 mg/seed, from about 0.01 to about 0.2 mg/seed, or from about0.02 to about 0.08 mg/seed.

The following examples are to be considered as merely illustrative, andare not intended to limit the scope of this invention.

EXAMPLES

Several active nematicidal compounds were combined with selecteddispersants and other excipients and used in preparation of suspensionconcentrate compositions in the following examples. The nematicidalcompounds are identified in Table 1.

TABLE 1 Ia-i 3-phenyl-5- (thiophen-2- yl)-1,2,4- oxadiazole

Ia-ii 3-(4- chlorophenyl)- 5-(furan-2-yl)- 1,2,4-oxadiazole

Ia-iii 3-(4-chloro-2- methylphenyl)- 5-(furan-2-yl)- 1,2,4-oxadiazole

Example 1: Preparation of a suspension concentrate comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)

A quantity of the nematicidal compound Ia-i (25.00 g) was added to anaqueous solution of water (25.00 g), glycerin (2.15 g), MORWET D-500dispersant (0.32 g), and AGNIQUE DF 6889 antifoam agent (0.05 g). Theresulting mixture was milled with a SIZEGVARI ATTRITOR milling systemmade by UNION PROCESS containing stainless steel beads having a diameterof ⅛ inch in a 100 mL jacketed metal container. The stirring speed wascontrolled by a VARIAC variable autotransformer.

After milling the mixture for 1 hour 40 minutes at a speed of 50 v/140v, a white aqueous suspension (45.25 g) was collected. The particle sizecharacteristics of the suspension were analyzed with a BECKMAN COULTERLS Particle Size Analyzer (model LS 13 320). The results indicated amean particle size of 4.896 μm, with a median particle size of 2.937 μm.The suspension was determined to contain 47.6% (w/w) of the Ia-inematicide.

Example 2: Preparation of a Suspension Concentrate Comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)

A quantity of the nematicidal compound Ia-i (30.00 g) was added to anaqueous solution of water (25.00 g), glycerin (3.00 g), MORWET D-500dispersant (0.60 g), and AGNIQUE DF 6889 antifoam agent (0.05 g). Theresulting mixture was milled with a SIZEGVARI ATTRITOR milling systemmade by UNION PROCESS containing stainless steel beads having a diameterof ⅛ inch in a 100 mL jacketed metal container. The stirring speed wascontrolled by a VARIAC variable autotransformer.

After milling the mixture for 1 hour 30 minutes at a speed of 50 v/140v, and an additional 2 hours 15 minutes at 40 v/140 v, a white aqueoussuspension (45.20 g) was collected. The suspension was determined tocontain 51.2% (w/w) of the Ia-i nematicide.

Example 3: Preparation of a suspension concentrate comprising3-(4-chlorophenyl)-5-(furan-2-yl)-1,2,4-oxadiazole (Ia-ii)

A quantity of the nematicidal compound Ia-ii (34.00 g) was added to anaqueous solution of water (25.00 g), glycerin (3.00 g), MORWET D-500dispersant (0.60 g), and AGNIQUE DF 6889 antifoam agent (0.10 g). Theresulting mixture was milled with a SIZEGVARI ATTRITOR milling systemmade by UNION PROCESS containing stainless steel beads having a diameterof ⅛ inch in a 100 mL jacketed metal container. The stirring speed wascontrolled by a VARIAC variable autotransformer.

After milling the mixture for 4 hours at a speed of 50 v/140 v, a whiteaqueous suspension (45.40 g) was collected. The particle sizecharacteristics of the suspension were analyzed with a BECKMAN COULTERLS Particle Size Analyzer (model LS 13 320). The results indicated amean particle size of 4.58 μm, with a median particle size of 3.14 μm.The suspension was determined to contain 54.2% (w/w) of the Ia-iinematicide.

Example 4: Preparation of a Suspension Concentrate Comprising3-(4-chloro-2-methylphenyl)-5-(furan-2-yl)-1,2,4-oxadiazole (Ia-iii)

A quantity of the nematicidal compound Ia-iii (34.00 g) was added to anaqueous solution of water (25.00 g), glycerin (3.00 g), MORWET D-500dispersant (0.60 g), and AGNIQUE DF 6889 antifoam agent (0.05 g). Theresulting mixture was milled with a SIZEGVARI ATTRITOR milling systemmade by UNION PROCESS containing stainless steel beads having a diameterof ⅛ inch in a 100 mL jacketed metal container. The stirring speed wascontrolled by a VARIAC variable autotransformer.

After milling the mixture for 4 hours at a speed of 50 v/140 v, a whiteaqueous suspension (49.10 g) was collected. The particle sizecharacteristics of the suspension were analyzed with a BECKMAN COULTERLS Particle Size Analyzer (model LS 13 320). The results indicated amean particle size of 3.217 μm, with a median particle size of 2.192 μm.The suspension was determined to contain 54.2% (w/w) of the Ia-iiinematicide.

Example 5: Preparation of a Suspension Concentrate Comprising3-(4-chlorophenyl)-5-(furan-2-yl)-1,2,4-oxadiazole (Ia-ii)

A quantity of the nematicidal compound Ia-ii (34.00 g) was added to anaqueous solution of water (141.67 g), glycerin (17.00 g), and MORWETD-500 dispersant (3.40 g). The resulting mixture was milled with aSIZEGVARI ATTRITOR milling system made by UNION PROCESS containingstainless steel beads having a diameter of ⅛ inch in a 500 mL jacketedmetal container. The stirring speed was controlled by a VARIAC variableautotransformer.

After milling the mixture for 1 hour at a speed of 75 v/140 v, a smallamount of AGNIQUE DF 6889 antifoam agent (0.10 g) was added. The mixturewas then further stirred at 75 v/140 v for 45 minutes, and at 60 v/140 vfor an additional 1 hour 45 minutes.

Following the milling process, a white aqueous suspension (330.5 g) wascollected from the container. The particle size characteristics of thesuspension were analyzed with a BECKMAN COULTER LS Particle SizeAnalyzer (model LS 13 320). The results indicated a mean particle sizeof 2.90 μm, with a median particle size of 1.74 μm. The suspension wasdetermined to contain 52.8% (w/w) of the Ia-ii nematicide.

Example 6: Preparation of a Suspension Concentrate Comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)

A quantity of the nematicidal compound Ia-i (34.00 g) was added to anaqueous solution of water (141.67 g), glycerin (17.00 g), and MORWETD-500 dispersant (3.40 g). The resulting mixture was milled with aSIZEGVARI ATTRITOR milling system made by UNION PROCESS containingstainless steel beads having a diameter of ⅛ inch in a 500 mL jacketedmetal container. The stirring speed was controlled by a VARIAC variableautotransformer.

After milling the mixture for 1 hour at a speed of 75 v/140 v, a smallamount of AGNIQUE DF 6889 antifoam agent (0.10 g) was added. The mixturewas then further milled at 75 v/140 v for 45 minutes and at 60 v/140 vfor an additional 1 hour 45 minutes.

Following the milling process, a white aqueous suspension (305.3 g) wascollected from the container. The particle size characteristics of thesuspension were analyzed with a BECKMAN COULTER LS Particle SizeAnalyzer (model LS 13 320). The results indicated a mean particle sizeof 3.334 μm, with a median particle size of 2.071 μm. The suspension wasdetermined to contain 52.8% (w/w) of the Ia-i nematicide.

Example 7: Effect of Milling Time on the Mean/Median Particle SizeDiameter of a Suspension Concentrate Comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)

A quantity of the nematicidal compound Ia-i (362.4 g) was added to anaqueous solution of water (283.34 g), glycerin (34.00 g), and MORWETD-500 dispersant (6.80 g). The resulting mixture was pre-milled with adissolver apparatus at 1900 rpm for 20 minutes. A portion of theresulting pre-milled slurry (60% of the total volume) was added to aNETZSCH MINI ZETA II milling machine filled with zirconium beads havinga diameter of 1.6-2 mm. The slurry was milled for 1 hour, after which asample of the resulting white slurry (250 g) was collected.

During the milling process, samples were periodically extracted foranalysis using a BECKMAN COULTER LS Particle Size Analyzer (model LS 13320). The resulting mean and median particle diameters for each sampleare summarized in Table 2 below:

TABLE 2 Milling Time (mins) Mean (μm) Median (μm) Mean/Median 15 4.0732.834 1.437 30 3.041 2.062 1.475 45 2.872 1.851 1.551 60 2.781 1.7601.580

The final suspension was determined to contain 44.2% (w/w) of the Ia-inematicide. This example demonstrates that the mean and/or medianparticle size of the formulation can be controlled as a function of thetotal milling time.

Example 8: Preparation of Seed Treatment Compositions

Seed treatment compositions were prepared using the suspensionconcentrate compositions prepared in Examples 2-4 above.

Composition 1: A seed treatment composition comprising the nematicidalcompound Ia-i was prepared by mixing a portion of the compositionprepared in Example 2 (8.00 g) with CF CLEAR seed coat polymer (0.30 g),BECKER-UNDERWOOD seed gloss (1.00 g), and BECKER-UNDERWOOD red colorcoating (2.00 g).

Composition 2: A seed treatment composition comprising the nematicidalcompound Ia-iii was prepared by mixing a portion of the compositionprepared in Example 3 (18.40 g) with CF CLEAR seed coat polymer (0.69g), BECKER-UNDERWOOD seed gloss (2.30 g), and BECKER-UNDERWOOD red colorcoating (4.60 g).

Composition 3: A seed treatment composition comprising the nematicidalcompound Ia-ii was prepared by mixing a portion of the compositionprepared in Example 4 (18.40 g) with CF CLEAR seed coat polymer (0.69g), BECKER-UNDERWOOD seed gloss (2.30 g), and BECKER-UNDERWOOD red colorcoating (4.60 g).

Example 9: Treatment of Seeds with Nematicidal Compositions

Soybean seeds (2.2 kg) were added to a WILLY NIKLAUS GMBH seed treatingapparatus. The seeds were tumbled inside the treater while a quantity ofseed treatment formulation was added. To ensure full dispersion of thetreatment composition, seeds were allowed to tumble for an additional 30seconds before being collected.

The amount of seed treatment composition used in each prepared samplewas varied in accordance with the targeted amount of active ingredientper seed. As shown in the table below, the targeted amount ranged from0.1 to 0.5 mg/seed for Ia-i, and from 0.1 to 1 mg/seed for Ia-iii andIa-ii. The actual amount of active ingredient per seed was analyzed uponremoval from the seed treatment apparatus. The results are summarized inthe table below, where the “Composition No.” refers to the compositions1-3 prepared in Example 8.

TABLE 3 Targeted Actual Amount of Composition Active Active LoadingActive Loading Composition No. Ingredient (mg/seed) (mg/seed) (g) 1 Ia-i0.1 0.07 0.98 1 Ia-i 0.3 0.22 2.94 1 Ia-i 0.5 0.37 4.90 3 Ia-ii 0.1 0.070.92 3 Ia-ii 0.3 0.25 2.77 3 Ia-ii 0.5 0.46 4.62 3 Ia-ii 0.0 0.83 9.24 2Ia-iii 0.1 0.04 0.92 2 Ia-iii 0.3 0.21 2.77 2 Ia-iii 0.5 0.40 4.62 2Ia-iii 0.0 0.65 9.24

The results indicate that, for each sample, a significant portion of theactive nematicidal ingredient added to the seed treatment apparatus wassuccessfully transferred to the seed.

Example 10: Preparation of Suspension Concentrate Compositions

An additional series of suspension concentrate compositions wereprepared using the procedures set forth below.

A stock buffer solution was prepared by adding anhydrous monobasicpotassium phosphate (9.361 g) and dibasic sodium phosphate heptahydrate(32.732 g) to a 1 liter volumetric flask, the balance of which wasfilled with deionized water. The flask was shaken until the salts werefully dissolved, providing a clear buffer solution with a pH of 7.

For each sample, a blank solution was then prepared by combining MORWETD-425 dispersant, PLURONIC L-35 secondary dispersant, propylene glycol,and a quantity of the stock buffer solution as prepared above. Therelative proportions of these components in each sample, respectively,are provided in Table 4 below.

In the next step of the process, the blank solution was mixed with aquantity of Ia-i nematicide and a small amount of BYK-016 antifoam agentin a 1 liter beaker. The formulation was then agitated with a Tekmarhomogenizer at 9,000 rpm for 10 to 12 minutes, resulting in a slurry.The particle size of the pre-milled slurry was measured with a BECKMANCOULTER LS Particle Size Analyzer (model LS 13 320).

For formulation Sample A and Sample C the pre-milled slurry was thenadded to a NETZSCH MINI ZETA II apparatus filled with either glass orzirconium oxide beads (200 mL) equipped with cooling water. Aftermilling for 35 minutes, the resulting white slurry was collected, andthe particle size was measured as described above. Formulation Sample Bwas pre-milled only to give a median particle size of 5.8 μm. Theparticle size can be reduced further through optimization of thepre-milling process.

A stabilizer composition was prepared by adding KELZAN CC stabilizingagent (4.00 g) and PROXEL GXL biocide (8.00 g) to deionized water(388.00 g). After agitation with a mechanical stirrer at roomtemperature for 30 minutes, a homogeneous viscous liquid was obtained.

The milled slurry was then mixed with a stabilizer composition in a 9:1weight ratio to provide a flowable suspension concentrate composition. Asummary of three representative composition samples prepared accordingto this process is provided below:

TABLE 4 Sample A Sample B Sample C Ingredient (wt. %) (wt. %) (wt. %)Ia-i 45.91 45.91 45.91 MORWET D-425 1.13 1.13 4.52 Propylene glycol 5.655.65 5.65 Water 35.99 35.99 32.60 BYK-016 0.31 0.31 0.31 PLURONIC ® L-350.06 0.06 0.06 Buffer solution 0.94 0.94 0.94 Stabilizer (1% solution)10.00 10.00 10.00

As indicated above, the compositions prepared according to this processwere all able to achieve an active ingredient loading of at least about45% by weight. Each of the compositions was measured to have an averagemedian particle size of from 1.0 to 1.2 microns, with a polydispersityindex (median/mean) of from 1.4 to 1.5. Each of the compositions wasobserved to be storage stable at room temperature for more than threemonths.

The formulations can also be prepared with Netzch Mini Zeta II millingmachine via a pass mode. In a typical example, the formulation was firstpre-milled with a homogenizer and then added to the milling machine.After the formulation was passed through the milling machine, it wascollected and then added to the milling machine again. After passingthrough the milling machine at 3504 rpm three times, the formulation wascollected and mixed with the KELZAN stabilizer composition to give thefinal formulation. The particle size of the formulation was measuredbefore the stabilizer was added. The formulations prepared by themultiple pass mode are shown in Table 5. The particle sizes for theseformulations are shown in Table 6.

TABLE 5 Sample D Sample E Sample F Ingredient (wt. %) (wt. %) (wt. %)Ia-i 47.79 47.79 47.79 MORWET D-425 2.26 2.26 2.26 ISOPAR M 2.26 2.26 —humic acid, sodium salt 2.26 — 2.26 Propylene glycol 5.65 5.65 5.65Water 39.06 41.32 41.32 BYK-016 0.31 0.31 0.31 PLURONIC ® L-35 0.06 0.060.06 Buffer solution 0.039 0.039 0.039 Stabilizer composition 0.10 0.100.10 1,2-benzisothiazolin-3-one 0.20 0.20 0.20

TABLE 6 Formulation Mean (μm) Median (μm) Mean/Median Sample D 2.63 1.871.41 Sample E 2.80 1.93 1.45 Sample F 2.37 1.62 1.46

Example 11: Differential Scanning Calorimetry Analysis

Eleven batches of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)were characterized for polymorphic form using differential scanningcalorimetry (DSC) analysis. DSC data were collected using a TAINSTRUMENTS Q2000 DSC apparatus.

For each batch, samples in the mass range of 1 to 10 mg were crimped inaluminum sample pans and scanned over a range of 25° C. to about 120°C., increasing at a rate of 2° C. to 10° C. per minute, and using anitrogen purge at 50 mL/min.

The melting point onset ranged from approximately 106° C. to 108° C.,with enthalpy of fusion ranging from approximately 108 to 122 J/g. Theresults are shown below in Table 7. Enthalpy of fusion measurements wereobtained on single sample analysis using a relatively small sample sizeof approximately 2 mg.

TABLE 7 DSC Analysis Summary Batch Melting Point Onset Enthalpy ofFusion (J/g) A 107.0 C. 116.6 B 107.7 C. 117.1 C 107.3 C. 118.9 D 107.0C. 119.4 E 107.4 C. 110.1 F 107.7 C. 121.7 G 107.0 C. 118.9 H 106.1 C.107.5 I 106.7 C. 110.0 J 107.3 C. 108.7 K 107.9 C. 111.0

The thermal behavior of batch G was determined using differentialscanning calorimetry and thermogravimetric analysis. The DSC thermogramexhibited a sharp melting endotherm with an onset of 106.9° C. and anenthalpy of fusion of 118.9 J/g.

Microscopic evaluation of lot G showed birefringent acicular to columnarshaped particles, ranging in size from approximately 5 to 100 microns.FIG. 1 shows the representative photomicrograph.

Example 12: Solvent Recrystallization

To perform the solvent-based portion of the polymorph screen, the3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole test material wasrecrystallized using various solvents under approximately 240 differentcrystal growth conditions. The scale of the recrystallizationexperiments was from approximately 0.5 mL to 15 ml. The crystal growthconditions were changed by using binary gradient arrays of solventmixtures and by changing the saturation temperature, growth temperatureand evaporation rate (rate of supersaturation generation).

Saturated solutions were prepared by agitating excess (as possible) testmaterial in contact with the various solvent systems at the saturationtemperature. If solids did not completely dissolve in the solvent, themother liquor was separated from the residual solids by filtration. Themother liquor was then heated above the saturation temperature(overheated) to dissolve any remaining solids. The temperature of eachsolution was then adjusted to the growth temperature and a controllednitrogen shear flow was introduced to begin solvent evaporation.

The recrystallization conditions for the seven solvent based panels usedduring the study are summarized in Table 8A. Each recrystallizationpanel contained from 27 to 96 wells. The wells within each panelcontained different solvent compositions. Because of the differentsolvent composition in each well, each well acted as a different crystalgrowth experiment. The compositional solvent matrices for the fiverecrystallization panels used during the solvent-based portion of thepolymorph screen are shown below in Tables 8B through 8F, respectively.Based on the XRD analysis carried out on the screening samples (seeExample 18, below) a new polymorph of3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole was discovered in theseexperiments. The starting material was designated as Form I, while thenew polymorph was designated as Form II.

TABLE 8A Summary of Recrystallization Panels N₂ Saturation OverheatGrowth Flow No. of Scale Temp. Temp. Temp. Rate Panel Wells (mL) Solvent(° C.) (° C.) (° C.) (psi) 1 34 15 Single/ 25 55 25 1.5 Binary 2 34 15Single/ 25 NA 80 1.5 Binary 4 27 15 Binary 25 55 50 1.5 6 27 15 Binary25 NA 65 1.5 7 96 0.5 Binary 25 50 40 2

TABLE 8B Recrystallization Panel 1 (Evaporated at Room Temp) WellSolvent Sample ID XRD Form 1 methanol RC1-1 Form I 2 ethanol RC1-2 FormI 3 trifluoroethanol RC1-3 Form I 4 1-propanol RC1-4 Form I 5 2-propanolRC1-5 Form I 6 1-butanol RC1-6 Form I 7 2-butanol RC1-7 Form I 8 waterRC1-8 NA 9 dimethyl formamide RC1-9 Form I 10 dimethylacetamide RC1-10Form I 11 butyl amine RC1-11 Form I 12 diisopropyl amine RC1-12 Form I13 pyridine RC1-13 Form I 14 nitromethane RC1-14 Form I 15 acetoneRC1-15 Form I 16 methyl ethyl ketone RC1-16 Form I 17 isopropyl etherRC1-17 Form I 18 Ethyl acetate RC1-18 Form I 19 methyl tert butyl etherRC1-19 Form I 20 isopropyl acetate RC1-20 Form I 21 tetrahydrofuranRC1-21 Form I 22 acetonitrile RC1-22 Form I 23 methylene chloride RC1-23Form I 24 chloroform RC1-24 Form I 25 toluene RC1-25 Form I 26 heptaneRC1-26 Form I 27 1,4 dioxane RC1-27 Form I 28 NMP RC1-28 NA/T 29 DMSORC1-29 NA/T 30 xylene RC1-30 Form I 31 butyl acetate RC1-31 Form I 322-methyl tetrahydrofuran RC1-32 Form I 33 propylene glycol RC1-33 NA/T34 glycerol/pyridine (2:13) RC1-34 NA/T

TABLE 8C Recrystallization Panel 2 (Evaporated at 80° C.) Well SolventSample ID XRD Form 1 methanol RC2-1 Form I + II 2 ethanol RC2-2 Form I 3trifluoroethanol RC2-3 Form I 4 1-propanol RC2-4 Form I II 5 2-propanolRC2-5 Form I 6 1-butanol RC2-6 Form I 7 2-butanol RC2-7 Form I + II 8water/acetone (7.5/7.5) RC2-8 Form I 9 DMF/1-butanol (7.5/7.5) RC2-9Form II 10 DMA/IPE (7.5/7.5) RC2-10 Form II 11 butyl amine RC2-11 Form I12 diisopropyl amine RC2-12 Form I + II 13 pyridine RC2-13 Form I 14nitromethane RC2-14 Form I + II 15 acetone RC2-15 Form I 16 methyl ethylketone RC2-16 Form II 17 isopropyl ether RC2-17 Form I 18 Ethyl acetateRC2-18 Form I + II 19 methyl tert butyl ether RC2-19 Form I 20 isopropylacetate RC2-20 Form I + II 21 tetrahydrofuran RC2-21 Form I 22acetonitrile RC2-22 Form I + II 23 methylene chloride RC2-23 Form I + II24 chloroform RC2-24 Form I 25 toluene RC2-25 Form I + II 26 heptaneRC2-26 Form I + II 27 1,4 dioxane RC2-27 Form I + II 28 NMP/MeOH(7.5/7.5) RC2-28 Form II 29 DMSO/EtOH (7.5/7.5) RC2-29 Form I 30 xyleneRC2-30 Form I 31 butyl acetate RC2-31 Form I + II 32 2-methyltetrahydrofuran RC2-32 Form I 33 PropGly/CHCl3 (7.5/7.5) RC2-33 Form I34 glycerol/pyridine (1:14) RC2-34 Form I

TABLE 8D Recrystallization Panel 4 (Evaporated at 50° C.) Solvent Matrixand XRD Result for Recrystallization Panel 4 Sample Ratio of SolventsCo/Anti- Solvent ID 1 2 3 Solvent DMF A 12:3 7.5:7.5 3:12 1-butanol DMAB 12:3 7.5:7.5 3:12 IPE MEK C 12:3 7.5:7.5 3:12 EtOH NMP D 12:3 7.5:7.53:12 MeOH TFE E 12:3 7.5:7.5 3:12 Water Xylene F 12:3 7.5:7.5 3:12 IPAEtOAc G 12:3 7.5:7.5 3:12 2-butanol 1,4 dioxane H 12:3 7.5:7.5 3:12Heptane DCM I 12:3 7.5:7.5 3:12 Acetonitrile Sample XRD Form Co/Anti-Solvent ID 1 2 3 Solvent 5 A Form I Form Form 1-butanol I + II I + IIDMA B Form I Form II Form IPE I + II MEK C Form Form I Form I EtOH I +II NMP D Form II Form I Form I MeOH TFE E Form II Form I No sample WaterXylene F Form I Form I Form I IPA EtOAc G Form I Form I Form I 2-butanol1,4 dioxane H Form I Form I Form I Heptane DCM I Form I Form I Form IAcetonitrile

TABLE 8E Recrystallization Panel 6 (Evaporated at 65° C.) Solvent Matrixand XRD Result for Recrystallization Panel 6 Sample Ratio of SolventsCo/Anti- Solvent ID 1 2 3 Solvent TFE A 12:3 7.5:7.5 3:12 IsopropylAcetate 1-propanol B 12:3 7.5:7.5 3:12 MEK THF C 12:3 7.5:7.5 3:12Chloroform Butylamine D 12:3 7.5:7.5 3:12 Toluene Diisopropyl- E 12:37.5:7.5 3:12 butyl amine acetate Pyridine F 12:3 7.5:7.5 3:12 2-meth THFNitromethane G 12:3 7.5:7.5 3:12 DMA Acetone H 12:3 7.5:7.5 3:12 NMPMTBE I 12:3 7.5:7.5 3:12 DMF Sample XRD Form Co/Anti- Solvent ID 1 2 3Solvent TFE A Form Form II Form Isopropyl I + II I + II Acetate1-propanol B Form Form Form I MEK I + II I + II THF C Form Form I Form IChloroform I + II Butylamine D Form I Form I Form I Toluene Diisopropyl-E Form I Form Form I butyl amine I + II acetate Pyridine F Form Form IForm I 2-meth THF I + II Nitromethane G Form Form I Form I DMA I + IIAcetone H Form Form I Amorphous/ NMP I + II LC MTBE I Form I Form I FormI DMF

TABLE 8F Recrystallization Panel 7 (96 Well Plate, Evaporated at 40° C.)Nitro- Isopropyl 1,4 Pyridine methane Acetone MEK EtOAc MTBE acetate THFDCM CHCl3 Toluene dioxane 1 2 3 4 5 6 7 8 9 10 11 12 A TFE Form I Form IForm Form Form I LC NA NA Form I NA Form Form I II II II B 1- NA FormI + Form NA Form I Form I Form I NA NA LC Form Form I propanol II II IIC IPA NA Form Form NA Form NA Form I NA NA NA NA NA II II II D 2- LCForm Form NA Form I NA Form I NA NA NA NA NA butanol II II E DMF NA NANA Form NA NA NA Form I Form NA NA NA II II F DMA NA NA NA NA Form I NANA NA NA NA Form I NA G butyl- NA Form NA NA NA Form I NA NA Form I NANA NA amine II H Di- Form Form I Form I NA NA NA NA NA NA NA NA NAisopropyl II amine

Example 13: Recrystallization from the Melt

Cyclic DSC analysis was performed on lot G (Form I) to determine if3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole would recrystallize from themelt as a different form (solvent-less recrystallization). Experimentswere performed by heating the material above the melting temperature,then cooling the material at a rate of 5° C., 10° C., 20° C., 30° C.,40° C. or 50° C. per minute, followed by reheating above the meltingtemperature. At the 5° C. to 30° C. per minute cooling rates, the firstenthalpy of fusion values (for the starting material) were approximately120 J/g while the second values (for the melting of the solids obtainedafter cooling the original melt) were approximately 100 J/g. There wasalso a slight change in the melting point onset (approximately 0.5° C.).It is believed that melting Form I, followed by recrystallization, mayresult in the formation of Form II.

The results of the experiments performed at cooling rates of 40° C. and50° C. per minute were unclear, and may indicate that the experiment wasuncontrolled under these conditions.

FIG. 3 shows a sample cyclic DSC thermogram from the run conducted at acooling rate of 30° C. per minute.

In a further experiment, approximately 300-400 mg of Form I startingmaterial was heated to melting in a forced air oven at approximately120° C. for approximately 40 minutes. The sample was slow cooled to roomtemperature, and XRD, DSC and proton NMR analyses were performed on thissample. The XRD pattern was different from the starting material (FormI) and was similar to the Form II pattern. DSC exhibited a melting onsettemperature of 107.8° C. and enthalpy of fusion of 103.2 J/g.

Example 14: Grinding Analysis

Batches of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole polymorphic FormsI and II were ground using a CRESCENT WIG-L-BUG ball mill for 2 minutesat 4800 oscillations per minute (3.2 m/s) in two separate experiments.Under these conditions, no transformation was observed in Form I, whilethe Form II sample transformed to Form I. FIG. 4 shows the XRD overlayof the milled Form I and Form II samples and the reference patterns ofForms I and II. The Form II used in this experiment was obtained byrecrystallization from the melt of Form I, as described in Example 14,above,

Example 15: Mechanical Pressure Analysis

Batches of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole polymorphic FormsI and II were placed in a CARVER press and compressed at approximately15,000 psi for approximately 20 seconds in two separate experiments. XRDanalysis was performed on the samples. The resulting XRD pattern matchedthe starting material in both experiments, as shown in FIGS. 5A and 5Bfor Forms I and II, respectively. The pressurized treatment did notreveal any changes in the polymorphic form of the starting material inboth experiments. The Form II used in this experiment was obtained byrecrystallization from the melt of Form I, as described in Example 14,above.

Example 16: Non-Competitive Slurry Experiments

In addition to the solvent recrystallization experiments,non-competitive slurry experiments were performed to search for newsolid-state forms of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole. Theseexperiments rely on solubility differences of different polymorphicforms (if the compound exists in different polymorphic forms). As such,only polymorphs having a lower solubility (that is, are more stable)than the original crystalline form can result from a noncompetitiveslurry experiment.

Essentially, when a solid is mixed with solvent to create slurry, asaturated solution eventually results. The solution is saturated withrespect to the polymorphic form dissolved. However, the solution issupersaturated with respect to any polymorphic form that is more stable(more stable forms have lower solubility) than the polymorphic forminitially dissolved. Therefore, any of the more stable polymorphic formscan nucleate and precipitate from solution. In addition, noncompetitiveslurry experiments are often useful in identifying solvents that formsolvates with the compound.

The slurry experiments were performed by exposing excess suppliedmaterial to solvents and agitating the resulting suspensions for severaldays at ambient temperature. The solids were filtered using a WHATMANGrade 1 apparatus (11 μm pore size) and analyzed by XRD to determine theresulting form(s). To avoid possible desolvation or physical changeafter isolation, the samples were not dried before X-ray analysis. Asummary of non-competitive slurry experiments is shown in Table 9.

TABLE 9 Vehicle Initial Form Duration Final Form Methanol I 12 days IEthanol I 12 days I Trifluoroethanol I 12 days I 1-propanol I 12 days IIsopropyl alcohol I 12 days I 1-butanol I 12 days I 2-butanol I 12 daysI water I 12 days I heptane I 12 days I glycerol/water (1:10) I 12 daysI propylene glycol/water (1:10) I 12 days I Isopropyl alcohol/water(1:1) I 12 days I ethanol II  7 days I trifluoroethanol II  7 days I1-propanol II  7 days I Isopropyl alcohol II  7 days I 1-butanol II  7days I 2-butanol II  7 days I heptane II  7 days I glycerol/water (1:10)II  7 days I propylene glycol/water (1:10) II  7 days I Isopropylalcohol/water(1:1) II  7 days I

Based on their X-ray scattering behavior, the slurry experiments withForm I as the starting material resulted in Form I after approximately12 days of slurring (indicating no transformation). The slurryexperiments with Form II as the starting material (obtained byrecrystallization from the melt, as set forth in Example 14, above)resulted in Form I after approximately 7 days of slurring. These dataindicate that Form I is more stable than Form II at ambient temperatureand pressure. No new polymorphs, solvates, or hydrates were isolated inthese experiments.

Example 17: X-Ray Analysis of Screening Samples

Batches of solid 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole polymorphsgenerated from the solvent based recrystallization panels and from othermeans (slurry, recrystallization from melt in an oven, etc.) wereanalyzed by powder XRD. To mitigate preferred grain effects, a twodimensional detection system was used to collect all the XRD screeningdata. The two dimensional detector integrates along the concentric Debyecones which helps reduce pattern variation. An example of the Debye coneintegration using a two dimensional detector is shown below. If brightspots appear in the conical rings, it indicates strong preferred graineffects that can lead to considerable variability in the observeddiffraction patterns including changes in peak intensities. Some samplesof 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole exhibited preferred graineffects based on the appearance of the scattering behavior.

The results of this analysis revealed the material exists as twodifferent polymorphs. The polymorphs were designated as Forms I and II.A powder XRD analysis of the Form I polymorphs, corresponding to theinitial test samples, is set forth in FIG. 6. A powder XRD analysis ofthe Form II polymorphs is set forth in FIG. 7.

The initial test material was designated as Form I. The resulting formdesignation for each individual (solvent-based) recrystallizationexperiment is shown in Tables 7B through 7F, above.

Example 18: Summary of Formation of Forms I and II

A number of different crystallization conditions were used to producethe samples utilized in Examples 12 through 18, above. Polymorphic FormI of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole was obtained inapproximately 50% of the experiments under various crystallizationconditions. Polymorphic Form II was obtained in approximately 10% of theexperiments also under various crystallization conditions. Mixtures ofForms I and II were obtained in approximately 11% of the experimentsindicating that the two polymorphs have a tendency to nucleate and growconcomitantly. Form I appears to be the thermodynamically stable formunder ambient conditions based on the results of the non competitiveslurry experiment. The exact crystallization conditions are shown inTables 7A through 7F, above.

Table 10 shows a summary of the results obtained in all the experimentpanels in this study. Note that Panels 1, 2, 4, 6, and 7 are describedin Example 13, above. Panel 3 corresponds to the recrystallization fromthe melt as set forth in Example 14, above. Panels 5 and 8 correspond tothe noncompetitive slurry experiments conducted with respect to Form Iand Form II, respectively, in Example 17, above.

TABLE 10 Mix of No. of Forms I No Panel No. Experiments Form I Form IIand II Result Panel 1 34 29 0 0 5 Panel 2 34 17 4 13 0 Panel 3 (Melt) 50 3 0 2 Panel 4 27 19 3 4 1 Panel 5 Form 1 12 12 0 0 0 NC Slurry Panel 627 16 1 9 1 Panel 7 96 well 96 19 14 1 62 Panel 8 Form 2 10 10 0 0 0 NCSlurry Total 245 122 25 27 71 % of total 100% 50% 10% 11% 29%

Example 19: Competitive Slurry Experiments

In addition to the solvent recrystallization experiments, a competitiveslurry experiment was also performed to determine the most stablepolymorphic form of 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole. Theseexperiments rely on the solubility differences of different polymorphicforms. As such, only polymorphic forms (and solvates) having a lowersolubility (more stable) than the form initially dissolved can resultfrom a competitive slurry experiment.

Essentially, when a solid is dissolved in a (slurry) solvent, asaturated solution eventually results. The solution is saturated withrespect to the polymorphic form dissolved. However, the solution issupersaturated with respect to any polymorphic form that is more stable(more stable forms have lower solubility) than the polymorphic forminitially dissolved. Therefore, any of the more stable polymorphic formscan nucleate and precipitate from solution. In addition, competitiveslurry experiments are often useful in identifying solvents that formsolvates with the API.

The slurry experiment was performed by exposing excess material of FormsI and II to a small volume of neat solvent and agitating the resultingsuspensions for several days at ambient temperature. The solids werefiltered and analyzed by XRD to determine the resulting form. To avoidpossible desolvation or physical change after isolation, the sample wasnot dried before x-ray analysis. Table 11 shows the results of thecompetitive slurry experiment.

TABLE 11 Initial Forms Slurry Final Form (XRD) Solvent Duration (XRD) I& II Isopropyl alcohol 1 week I

The thermal data obtained above was used to calculate an approximatevalue for the transition temperature of conversion of Forms I and IIusing methods known in the art. The value obtained using this method wasapproximately 102° C. Based on these calculations, Form I is expected tobe the stable form below this temperature and Form II above it. This isanother characteristic of an enantiotropic polymorphic relationship.

A graphical XRD overlay of the competitive slurry experiment is depictedin FIG. 8.

Example 20: Estimation of Transition Temperature

Polymorphic Forms I and II of3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole, as well as a 50/50 mixturethereof, were analyzed by DSC at a slow heating rate of 2° C. perminute, with similar sample sizes. The melting temperatures and enthalpyof fusion data are shown in Table 12, below. These data indicate thatForm I has a lower melting temperature and a higher enthalpy of fusion.Form II has a higher melting temperature and a lower enthalpy of fusion.In accordance with the Heat of Fusion Rule, this indicates that Form Iand II have an enantiotropic relationship. FIGS. 9A through 9C show therelevant DSC thermograms for Form I, Form II, and a mixture of Forms Iand II, respectively.

The thermal data using the procedure set forth above was used tocalculate an approximate value for the transition temperature ofconversion of Forms I and II, resulting in an estimated transitiontemperature value of 102° C. Based on these calculations, Form I isexpected to be the stable form below this temperature, while Form II isexpected to possess greater thermodynamic stability above thattemperature. This further indicates that Forms I and II exhibit anenantiotropic polymorphic relationship.

TABLE 12 Onset Maximum Enthalpy of Fusion Sample ID (° C.) (° C.) (J/g)Batch G Form I 106.9 107.9 117.9 54478-21-4 Form II 108.0 108.8 98.350/50, Form I/II 108.0 108.0, 108.8 114.6

Example 21: Storage Stability of Polymorphs

To determine the storage stability and/or hydrate formation of3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole Form I material duringstorage at ambient conditions, samples were monitored in two statichumidity chambers. In these studies, samples were stored in open Petridishes in chambers containing saturated salt solutions to maintain therelative vapor pressure. Solutions of saturated potassium chloride (84%RH) and sodium chloride (75% RH) salts at ambient temperature were used.

FIG. 10 shows the XRD pattern of the samples stored at 75 and 84% RHafter 4 weeks of storage. As indicated in the figure, Form I does notform a hydrate and appears to be thermodynamically stable over time atambient conditions.

In contrast, samples of Form II stored in a scintillation vial in a hoodunder ambient conditions showed signs of transformation to Form I whenanalyzed by XRD after approximately 6 days of storage. FIG. 11 shows theXRD overlays of Forms I, II and the sample of Form II which showed signsof transformation to Form I.

Example 22: Soybean Cyst Nematode Assay

Formulations were tested for nematicidal activity against soybean cystnematode (SCN) in an SCN cup assay.

The formulations were prepared as follows:

Preparation of the phosphate buffer solution: To a 1 L volumetric flaskwere added potassium phosphate monobasic anhydrous (9.329 g) and sodiumphosphate dibasic heptahydrate (32.756 g). DI water was added to theflask to the mark and it was inverted 15 times to give a clear solution.

Preparation of Formulation Blank A:

To a 2 L beaker were added MORWET D-425 (43.6 g), DI water (1,386.9 g),the phosphate buffer solution (36.3 g), propylene glycol (217.7 g), andPLURONIC L-35 (2.2 g). The mixture was stirred with a spatula to give abrown solution.

Preparation of Formulation Blank B:

To a 2 L beaker were added MORWET D-425 (174.3 g), DI water (1.256.0 g),the phosphate buffer solution (36.3 g), propylene glycol (217.8 g), andPLURONIC L-35 (2.1 g). The mixture was stirred with a spatula to give adark brown solution.

Preparation of the KELZAN Stabilizer Solution:

To a 1 L beaker were added KELZAN CC (4.060 g), PROXEL GXL (7.978 g),and DI water (388.273 g). The mixture was then agitated with a Meltonmechanical stirrer (model CM-100) at 2,000 rpm for 30 minutes to give aviscous liquid.

Preparation of Suspension Concentrate Formulation 3:

To a 2 L beaker were added Formulation Blank A (497.3 g), Compound Ia-i(521.4 g), and BYK-016 (3.6 g). The mixture was stirred with a spatulato give a slurry. The mixture was placed in an ice bath and a TekmarT554 homogenizer (model TR-10) was used for the pre-milling. During thepre-milling, the slurry (1022.3 g) was agitated with the homogenizer at9,000 rpm for 12 mins. An Eiger mill (model M250) was filled withzirconium oxide beads with an average diameter of 0.3-0.4 mm. Nearlyhalf of the pre-milled slurry (501.4 g) was then added to the Eiger milland was milled with a speed of 5000 rpm in recycle mode at roomtemperature. After 30 minutes, the resulting white liquid formulation(412.4 g) was collected and mixed with the KELZAN stabilizer solution(45.8 g) to give the final formulation (458.2 g). The particle size ofthe formulation was analyzed with a Beckman Coulter particle sizeanalyzer (Model LS 13 320) before the stabilizer was added.

Preparation of Suspension Concentrate Formulation 4:

The pre-milled slurry (501.4 g) from the suspension concentrateformulation above was also milled with the same Eiger mill filled withzirconium oxide beads with an average diameter of 0.3-0.4 mm. Aftermilling for 120 minutes, the resulting white liquid formulation (408.5g) was collected and mixed with the KELZAN stabilizer solution (45.4 g)to give the final formulation (453.9 g). The particle size of theformulation was also analyzed with a Beckman Coulter particle sizeanalyzer (Model LS 13 320) before the stabilizer was added.

Preparation of Suspension Concentrate Formulation 5:

To a 1 L beaker were added Formulation Blank B (383.3 g), Compound Ia-i(261.1 g), and BYK-016 (2.5 g). The mixture was stirred with a spatulato give a slurry. The mixture was placed in an ice bath and a TekmarT554 homogenizer (model TR-10) was used for the pre-milling. During thepre-milling, the slurry was agitated with the homogenizer at 9,000 rpmfor 10 mins. The milling was divided into two stages. Both Netzsch MiniZeta II filled with glass beads with an average diameter of 0.8-1 mm andEiger mill (model M250) filled with zirconium oxide beads with anaverage diameter of 0.1-0.2 mm were used in the milling. In the firststage, the slurry was passed through the Netzsch miller three times andthe miller was operated at 3,504 rpm for each pass. In the second stage,the slurry was passed through the Eiger miller ten times and the millingwas operated at 5,000 rpm. A white liquid (452.1 g) was collected andpart of the white liquid (349.0 g) mixed with the KELZAN stabilizersolution (38.8 g) to give the final formulation (387.8 g). The particlesize of the formulation was also analyzed with a Beckman Coulterparticle size analyzer (Model LS 13 320) before the stabilizer wasadded.

Preparation of Suspension Concentrate Formulation 6:

To an 8 dram vial were added MORWET D-425 (0.714 g), DI water (3.75 g),the phosphate buffer solution (0.147 g), ISOPAR M (1.45 g), propyleneglycol (0.898 g), PLURONIC L-35 (0.009 g), Compound Ia-i (7.315 g), andBYK-016 (0.067 g). The mixture was stirred followed by addition of 3 mmdiameter stainless steel beads (14 mL). The vial was capped and placedon a US Stoneware roller (Ser. No. CK-11009) and rotated at a speedsetting of 50. After 2 days the slurry (5.903 g) was collected and mixedwith the KELZAN stabilizer solution (0.660 g) to give the finalformulation (6.563 g). The particle size of the formulation was analyzedwith a Beckman Coulter particle size analyzer (Model LS 13 320) beforethe stabilizer was added.

Table 13 below depicts the compositions of each formulation used forseed treatment in the SCN efficacy assay.

TABLE 13 Composition of Formulation for Seed Treatment Ia-i CommercialFormula- Formula- Seed Compound Treat- tion tion Treatment Water Ia-iRate ment Ia-i (g) (g) (g) (mg/seed) 1  NA N/A N/A N/A N/A 2  NA 0 1.5570.64 N/A 3A 3 0.36 0 0.64 0.05 3B 3 2.16 0 1.01 0.3 4A 3 0.36 1.557 0.640.05 4B 3 2.16 1.557 1.01 0.3 5A 4 0.36 0 0.64 0.05 5B 4 2.16 0 1.01 0.36A 4 0.36 1.557 0.64 0.05 6B 4 2.16 1.557 1.01 0.3 7A 5 0.45 0 1.21 0.057B 5 2.73 0 1.16 0.3 8A 5 0.45 1.557 1.21 0.05 8B 5 2.73 1.557 1.16 0.39A 6 0.36 0 0.64 0.05 9B 6 2.16 0 1.01 0.3 10A  6 0.36 1.557 0.64 0.0510B  6 2.16 1.557 1.01 0.3

SCN Efficacy Assay

A4630 soybean plants were grown in cups filled with full strengthMurashige & Skoog basal salts fertilizer (Phytotech Cat. No. 201080-52)followed by 180 ml of 20:80 soil/sand mixture (sterile St. Charles sandand US 10 soil premixed by Hummert). A Gustafson Batch Modular Coater(BMC) Treater was used to the treat the soybean seeds with theformulations as described in Table 13.

The untreated seed and treated seed were placed on top of 20:80 soil andpushed ½ inch deep into the soil. The cups were placed in the growthchamber and the soil was misted with water to saturation. Propagationdomes were placed over the cups until the seeds had germinated (about 3to 5 days). Conditions in the growth chamber were as follows: 28° C.,60% relative humidity, and 16 h/14 h day/night periods, with 347μEinsteins of light.

Ten days after planting, soybean cyst inoculum (2×500 μL, 5000 eggs/cup)was delivered into the soil on two sides of the soybean plant. Theplants were grown for an additional 5 weeks after inoculation andwatered as needed with overhead watering.

The efficacy of the formulations was determined by harvesting plants (45days) and counting cysts. Table 14 summarizes the bioefficacy againstSCN at 50 μg/seed and 300 μg/seed.

TABLE 14 Treat- Particle Rate Cyst Counts ment Size (μm) (mg/seed) MeanStd Dev Std Err Mean 1  N/A 227 159 65 2  N/A 337 205 84 3A 0.8 0.05 14980 33 3B 0.3 67 47 19 4A 0.8 0.05 247 244 100 4B 0.3 92 106 43 5A 0.480.05 146 55 22 5B 0.3 90 58 24 6A 0.48 0.05 203 193 79 6B 0.3 57 71 297A 0.065 0.05 137 86 35 7B 0.3 150 55 25 8A 0.065 0.05 176 101 41 8B 0.386 70 29 9A 1.7 0.05 147 97 40 9B 0.3 80 89 36 10A  1.7 0.05 92 63 2810B  0.3 76 64 26

Example 23: Preparation of Suspension Concentrates Comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i) and imidacloprid

Several suspension concentrate co-formulation compositions comprisingthe nematicidal compound 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole(Ia-i) and imidacloprid were prepared. A summary of five representativeco-formulation compositions prepared in this Example is provided belowin Tables 15-19.

The phosphate buffer was prepared by adding potassium phosphatemonobasic anhydrous (9.361 g) and sodium phosphate dibasic heptahydrate(32.732 g) to a 1 L volumetric flask. Then DI water was added to theflask to the mark. After it was shaken for a while, all the salts weredissolved to give a clear phosphate buffer with pH at 7.

The KELZAN thickener/stabilizer was prepared by adding KELZAN CC (4.00g) and PROXEL GXL (8.00 g) to DI water (388.00 g) and agitating with amechanical disperser at 2,900 rpm at room temperature for 30 minutes toobtain a viscous liquid (400 g).

The suspension concentrate co-formulation compositions were prepared bymixing all the ingredients in Part A at 300 rpm for 30 minutes or untilthe MORWET D-425 dissolved; adding all the ingredients in Part B to PartA and mixing at 300 rpm for 5 minutes; adding all the ingredients inPart C to the mixture of Parts A and B and homogenizing the solution at9,000 rpm for 10 minutes; preparing a combination of ground (grindingwas done using the roller with grinding media (Very High DensityZirconium Oxide Grinding Media, Yttria stabilized ½×½ cylinders)) Ia-iand imidacloprid (Part D) in a jar and pouring the solution above (PartsA, B, and C) in a controlled path; homogenizing the solution at 9000 rpmfor 30 minutes with an ice bath; milling the solution 4 times to achievethe desired particle size; adding Part E to the collected sample andmixing at 500 rpm for 10 minutes; and adding Parts F, G and H (ifpresent) to the collected sample and mixing at 900 rpm for 30 minutes.

ATLOX 4913 (available from Croda) is used as a crystal growth inhibitor.The weight percentage values reported in Tables 15-19 are theoretical,and were calculated based upon the weight of the actives added. Particlesizes reported in Tables 15-19 were measured by Beckman Coulter particlesize analyzer (Beckman Coulter LS 13 320 laser diffraction particle sizeanalyzer).

TABLE 15 Co-Formulation of Ia-i and Imidacloprid (particle size 2.196μm) Amount Theoretical Actual Part Ingredient (g) Wt % Wt % A MORWETD-425 0.36 2.17 DI water 2.45 14.78 B Buffer 0.15 0.90 DI water 2.4414.72 C Propylene glycol 0.90 5.43 ISOPAR M 0.36 2.17 Antifoam 0.05 0.30PLURONIC L-35 0.009 0.05 D Ia-i 5.93 35.77 34.1 Imidacloprid 1.68 10.138.7 E Stabilizer 1.59 9.60 Humic acid sodium salt 0.66 3.98

TABLE 16 Co-Formulation of Ia-i and Imidacloprid (particle size 1.91 μm)Amount Theoretical Actual Part Ingredient (g) Wt % Wt % A MORWET D-4250.36 2.17 DI water 2.45 14.74 B Buffer 0.15 0.90 DI water 2.44 14.68 CPropylene glycol 0.90 5.42 ISOPAR M 0.36 2.17 Antifoam 0.05 0.30PLURONIC L-35 0.009 0.05 D Ia-i 5.17 31.11 28.97 Imidacloprid 2.44 14.6813.17 E ATLOX 4913 0.34 2.05 F Stabilizer 1.59 9.58 G Humic acid sodiumsalt 0.36 2.17

TABLE 17 Co-Formulation of Ia-i and Imidacloprid (particle size 1.95 μm)Amount Theoretical Actual Part Ingredient (g) Wt % Wt % A MORWET D-4250.36 2.17 DI water 2.45 14.74 B Buffer 0.15 0.90 DI water 2.44 14.68 CPropylene glycol 0.90 5.42 ISOPAR M 0.36 2.17 Antifoam 0.05 0.30PLURONIC L-35 0.009 0.05 D Ia-i 5.17 31.11 28.82 Imidacloprid 2.44 14.6812.97 E ATLOX 4913 0 0 F Stabilizer 1.59 9.58 G Humic acid sodium salt 00.00 H Water 0.7 4.21

TABLE 18 Co-Formulation of Ia-i and Imidacloprid (particle size 1.94 μm)Amount Theoretical Actual Part Ingredient (g) Wt % Wt % A MORWET D-4250.36 2.17 DI water 2.45 14.74 B Buffer 0.15 0.90 DI water 2.44 14.68 CPropylene glycol 0.90 5.42 ISOPAR M 0.36 2.17 Antifoam 0.05 0.30PLURONIC L-35 0.009 0.05 D Ia-i 5.17 31.11 29.11 Imidacloprid 2.44 14.6813.19 E ATLOX 4913 0 0 F Stabilizer 1.59 9.58 G Humic acid sodium salt0.36 2.17 H Water 0.34 2.05

TABLE 19 Co-Formulation of Ia-i and Imidacloprid (particle size 1.92 μm)Amount Theoretical Actual Part Ingredient (g) Wt % Wt % A MORWET D-4250.36 2.17 DI water 2.45 14.74 B Buffer 0.15 0.90 DI water 2.44 14.68 CPropylene glycol 0.90 5.42 ISOPAR M 0.36 2.17 Antifoam 0.05 0.30PLURONIC L-35 0.009 0.05 D Ia-i 5.17 31.11 28.95 Imidacloprid 2.44 14.6813.36 E ATLOX 4913 0.34 2.05 F Stabilizer 1.59 9.58 G Humic acid sodiumsalt 0 0.00 H Water 0.36 2.17

Example 24: Aged Stability Studies of Suspension Concentrates Comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i) and imidacloprid

The five suspension concentrate co-formulation compositions comprisingthe nematicidal compound 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole(Ia-i) and imidacloprid prepared in Example 23 were subjected to an agedstability study.

In this study, particle size was measured by Beckman Coulter particlesize analyzer (Beckman Coulter LS 13 320 laser diffraction particle sizeanalyzer), viscosity was measured by Brookfield R/S plus Rheometer, andthe wt % values of Ia-i and imidacloprid were determined using HPLC.

Each concentrate co-formulation composition was sampled into individualjars and labeled for room temperature (RT) time 0, RT 4 weeks, RT 8weeks, 50° C. 4 weeks and 50° C. 8 weeks. Both 50° C. 4 weeks and 50° C.8 weeks samples were stored in a lab oven with temperature set at 50° C.

At time 0, RT time 0 samples were tested. At 4 weeks, the 50° C. 4 weekssamples were taken out of oven and set on the bench for 24 hours to letthe temperature reach RT. Then the RT 4 weeks samples and 50° C. 4 weekssamples were tested. At 8 weeks, the 50° C. 8 weeks samples were takenout of oven and set on the bench for 24 hours to let the temperaturereach RT. Then the RT 8 weeks samples and 50° C. 8 weeks samples weretested.

The testing results of the aged stability study for the five suspensionconcentrate co-formulation compositions summarized in Tables 15-19 arereported below in Tables 20-24, respectively. Note that there is aslight discrepancy in the measured concentrations of Ia-i andimidacloprid as compared to the theoretical values reported above inTables 15-19 for reasons known in the art; for example, the actives maynot have been 100% pure, or there may have been loss of active duringthe milling and/or processing of the sample.

TABLE 20 Aged Stability Studies for Composition in Table 15 Time 4 weeks4 weeks 8 weeks 8 weeks 0 (RT) (50° C.) (RT) (50° C.) Concentration Ia-i(wt %) 34.1 34.3 34.2 33.8 33.7 Concentration 8.7 8.88 9.08 8.8 8.6imidacloprid (wt %) Particle size (μm) 2.196 1.949 2.061 2.094 1.927Viscosity (cP) 134.68 185.35 212.85 164.97 202.32 pH 9.28 9.25 9.09 9.328.91

TABLE 21 Aged Stability Studies for Composition in Table 16 Time 4 weeks4 weeks 8 weeks 8 weeks 0 (RT) (50° C.) (RT) (50° C.) Concentration Ia-i(wt %) 28.97 29.43 29.43 30.13 30.12 Concentration 13.17 14.45 14.4213.9 13.86 imidacloprid (wt %) Particle size (μm) 1.913 1.97 1.972 1.861.904 Viscosity (cP) 92.46 101.54 123.32 99.82 119.71 pH 9.09 8.91 8.448.83 8.55

TABLE 22 Aged Stability Studies for Composition in Table 17 Time 4 weeks4 weeks 8 weeks 8 weeks 0 (RT) (50° C.) (RT) (50° C.) Concentration Ia-i(wt %) 28.82 29.49 28.95 30.1 30.09 Concentration 12.97 14.52 14.2113.98 13.95 imidacloprid (wt %) Particle size (μm) 1.95 1.917 1.9611.813 1.832 Viscosity (cP) 49.68 61.6 68.1 52.19 61.92 pH 8.55 8.36 8.398.22 8.28

TABLE 23 Aged Stability Studies for Composition in Table 18 Time 4 weeks4 weeks 8 weeks 8 weeks 0 (RT) (50° C.) (RT) (50° C.) Concentration Ia-i(wt %) 29.11 29.38 29.4 29.97 30.21 Concentration 13.19 14.37 14.1313.91 13.76 imidacloprid (wt %) Particle size (μm) 1.939 1.867 1.9241.817 1.883 Viscosity (cP) 77.03 86.7 104.39 82.64 101.28 pH 9.5 9.298.86 9.18 8.78

TABLE 24 Aged Stability Studies for Composition in Table 19 Time 4 weeks4 weeks 8 weeks 8 weeks 0 (RT) (50° C.) (RT) (50° C.) Concentration Ia-i(wt %) 28.95 29.34 29.22 29.73 30.12 Concentration 13.36 14.4 14.4 13.8913.99 imidacloprid (wt %) Particle size (μm) 1.916 1.902 1.885 1.8591.864 Viscosity (cP) 61.23 63.54 70.48 56.8 64.73 pH 7.36 7.29 7.3 7.167.21

Example 25: Preparation of Suspension Concentrates Comprising3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)

Several suspension concentrate formulation compositions comprising thenematicidal compound 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i)were prepared. A summary of three representative suspension concentrateformulation compositions prepared in this Example is provided below inTables 25-27.

The phosphate buffer was prepared by adding potassium phosphatemonobasic anhydrous (9.361 g) and sodium phosphate dibasic heptahydrate(32.732 g) to a 1 L volumetric flask. Then DI water was added to theflask to the mark. After it was shaken for a while, all the salts weredissolved to give a clear phosphate buffer with pH at 7.

The KELZAN thickener/stabilizer was prepared by adding KELZAN CC (4.00g) and PROXEL GXL (8.00 g) to DI water (388.00 g) and agitating with amechanical disperser at 2,900 rpm at room temperature for 30 minutes toobtain a viscous liquid (400 g). The same composition and preparationprocedure were used to prepare KELZAN thickener/stabilizer with a scaleup to 1 kg.

Ground Ia-i was prepared by adding it to a plastic bottle half filledwith cylindrical ceramic beads (½″ O.D.×½″ long) and rotated for 30minutes on a roller. Ground Ia-i was then collected after sievingthrough a screen and used in preparation of the suspension concentrateformulation.

The suspension concentrate formulation compositions were prepared byfirst preparing a formulation blank by adding MORWET D-425 (81.6 g) toDI water (1027.4 g) in a 1 gallon jar. After MORWET D-425 was dissolved,propylene glycol (204.1 g) phosphate buffer (34.0 g), PLURONIC L-35(2.04 g), antifoam (11.34 g), and ISOPAR M (81.6 g) were added to thejar and it was then agitated in an ice bath with a homogenizer at 9,000rpm for 6 minutes to give a brown emulsion (1442.1 g).

To a 1 gallon jar were added the formulation blank and ground Ia-i, andthe resulting mixture was well stirred with a spatula. Then the jar wasplaced in an ice bath and a Tekmar homogenizer (model T 45 S4) wasinserted into the slurry and the head of the homogenizer in the centerof the jar was about 5 mm above the bottom of the jar. Initially it wasoperated at 10,000 rpm for 3 minutes and then at 9,000 rpm for 27minutes. The particle size of the resulting slurry was measured after itwas milled for 30 minutes. If large particles still existed in theformulation slurry, it was milled for additional 5 to 10 minutes.Particle size was measured by Beckman Coulter particle size analyzer(Beckman Coulter LS 13 320 laser diffraction particle size analyzer).

A NETZCH MINI ZETA II mill apparatus filled with glass beads with adiameter of 0.7-1 mm (200 ml) was used in the milling. The mill wasconnected to the water line and cold water was used to control thetemperature increase during the milling. Before it was used, smallamount of the formulation blank (15.0 g) was added to the machine firstand the machine was then run at 3504 rpm for 30 s. Compressed nitrogenwas used to push the residual formulation blank out of the machine.

For the preparation of the suspension concentrate formulations in thisexample, the pass method was used to reduce the particle size of theformulation. During the milling, the formulation slurry obtained frompre-milling as described above was added to the mill when operated at3504 rpm. After milled through the mill, the formulation was collected.Then the same milling was repeated twice to give the formulation with anaverage median particle of about 2 μm with a particle size range of fromabout 1.6 to about 2.5 μm. If the particle size is still large, thefourth or additional milling is required.

In post treatment, the KELZAN thickener/stabilizer, humic acid sodiumsalt, and ATLOX 4913 were added to the collected formulation slurry andit was then stirred with a mechanical stirrer at room temperature for 30minutes to obtain the suspension concentrate formulation in the form ofa brown slurry. The wt % values reported in Tables 25-27 for the ATLOX4913, stabilizer, humic acid sodium salt, and water are based on sampleamounts recovered after milling (for example, if a total of 1000 g offormulation blank and nematicidal component are milled and 800 g arerecovered after milling, the amount of post milling components added arebased on 800 g).

TABLE 25 Formulation 7 Ingredient Wt % MORWET D-425 2.17 Buffer 0.91Propylene glycol 5.43 ISOPAR M 0.36 Antifoam (1520-US) 0.30 PLURONICL-35 0.05 Ia-i 45.88 ATLOX 4913 4.00 Stabilizer 9.60 Humic acid sodiumsalt 2.17 Water 4.53

TABLE 26 Formulation 8 Ingredient Wt % MORWET D-425 2.17 Buffer 0.91Propylene glycol 5.43 ISOPAR M 2.17 Antifoam (1520-US) 0.30 PLURONICL-35 0.05 Ia-i 45.88 ATLOX 4913 4.00 Stabilizer 9.60 Humic acid sodiumsalt 2.17 Water 27.32

TABLE 27 Formulation 9 Ingredient Wt % MORWET D-425 2.17 Buffer 0.91Propylene glycol 5.43 ISOPAR M 2.17 Antifoam (BYK-016) 0.30 PLURONICL-35 0.05 Ia-i 45.88 ATLOX 4913 4.00 Stabilizer 9.60 Humic acid sodiumsalt 2.17 Water 27.32

Example 26: Preparation of Suspension Concentrates ComprisingNematicidal Compound and a Second Active

Several suspension concentrate co-formulation compositions comprisingthe nematicidal compound 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole(Ia-i) or 3-(4-chloro-2-methylphenyl)-5-(furan-2-yl)-1,2,4-oxadiazole(Ia-iii) and various fungicides or insecticides as a second active wereprepared. A summary of sixteen representative co-formulationcompositions (A-P) prepared in this Example is provided below in Tables28-33.

The co-formulation compositions were prepared by adding DI water, MORWETD-425, phosphate buffer (as described above in Example 23) and propyleneglycol to a 30 ml vial equipped with cap. After MORWET D-425 wasdissolved, humic acid sodium salt, ISOPARM, antifoam (AGNIQUE DFM 111S),and PLURONIC L-35 were added to the vial and it formed a brown liquidafter stirring. Then the actives were added to the vial according to thecomposition in each formulation. Stainless steel beads (14 ml) with adiameter of 2 mm were added to the vial and the vial was capped tightly.The vial was placed in a large jar (16 oz.) and the large jar was caped.The large jar with four vials was placed on a roller (model 764 AVM fromU.S. Stoneware), and it was rolled at half of the maximum speed for 2days at ambient temperature. KELZAN thickener/stabilizer (as describedabove in Example 23) was then added to the vial and rolled at 10-20% ofthe maximum speed for 4 hrs. The formulation, which was flowable in thevial, was collected and a small amount of the sample was taken forparticle size analysis. The wt % values reported in Tables 28-33 aretheoretical based on the weight of the active added. The purities of themetalaxyl, tebuconozole and kresoxim methyl actives used in this Examplewere 90%, 96.8% and 97.5%, respectively.

TABLE 28 Co-Formulation of Ia-i and Imidacloprid A B C Amount AmountAmount Ingredient (g) Wt % (g) Wt % (g) Wt % MORWET D-425 0.36 2.26 0.362.26 0.36 2.26 Buffer 0.15 0.94 0.15 0.94 0.15 0.94 Propylene glycol0.90 5.66 0.90 5.66 0.90 5.66 ISOPAR M 0.36 2.26 0.36 2.26 0.36 2.26Antifoam 0.05 0.31 0.05 0.31 0.05 0.31 PLURONIC L-35 0.009 0.06 0.0090.06 0.009 0.06 Ia-i 3.80 23.88 0.76 4.77 6.85 43.03 Imidacloprid 3.8023.88 6.85 43.03 0.76 4.77 Humic acid sodium 0.36 2.26 0.36 2.26 0.362.26 salt Stabilizer 1.59 10.00 1.59 10.00 1.59 10.00 Water 4.53 28.474.53 28.47 4.53 28.47

TABLE 29 Co-Formulation of Ia-i and Tebuconazole D E F Amount AmountAmount Ingredient (g) Wt % (g) Wt % (g) Wt % MORWET D-425 0.36 2.26 0.362.26 0.36 2.26 Buffer 0.15 0.94 0.15 0.94 0.15 0.94 Propylene glycol0.90 5.66 0.90 5.66 0.90 5.66 ISOPAR M 0.36 2.26 0.36 2.26 0.36 2.26Antifoam 0.05 0.31 0.05 0.31 0.05 0.31 PLURONIC L-35 0.009 0.06 0.0090.06 0.009 0.06 Ia-i 3.80 23.88 0.76 4.77 6.85 43.03 Tebuconazole 3.8023.88 6.85 43.03 0.76 4.77 Humic acid 0.36 2.26 0.36 2.26 0.36 2.26sodium salt Stabilizer 1.59 10.00 1.59 10.00 1.59 10.00 Water 4.53 28.474.53 28.47 4.53 28.47

TABLE 30 Co-Formulation of Ia-i and kresoxim-methyl G H I Amount AmountAmount Ingredient (g) Wt % (g) Wt % (g) Wt % MORWET D-425 0.36 2.26 0.362.26 0.36 2.26 Buffer 0.15 0.94 0.15 0.94 0.15 0.94 Propylene glycol0.90 5.66 0.90 5.66 0.90 5.66 ISOPAR M 0.36 2.26 0.36 2.26 0.36 2.26Antifoam 0.05 0.31 0.05 0.31 0.05 0.31 PLURONIC L-35 0.009 0.06 0.0090.06 0.009 0.06 Ia-i 3.80 23.88 0.76 4.77 6.85 43.03 kresoxim-methyl3.80 23.88 6.85 43.03 0.76 4.77 Humic acid 0.36 2.26 0.36 2.26 0.36 2.26sodium salt Stabilizer 1.59 10.00 1.59 10.00 1.59 10.00 Water 4.53 28.474.53 28.47 4.53 28.47

TABLE 31 Co-Formulation of Ia-i and metalaxyl J K L Amount Amount AmountIngredient (g) Wt % (g) Wt % (g) Wt % MORWET D-425 0.36 2.26 0.36 2.260.36 2.26 Buffer 0.15 0.94 0.15 0.94 0.15 0.94 Propylene glycol 0.905.66 0.90 5.66 0.90 5.66 ISOPAR M 0.36 2.26 0.36 2.26 0.36 2.26 Antifoam0.05 0.31 0.05 0.31 0.05 0.31 PLURONIC L-35 0.009 0.06 0.009 0.06 0.0090.06 Ia-i 3.80 23.88 0.76 4.77 6.85 43.03 metalaxyl 3.80 23.88 6.8543.03 0.76 4.77 Humic acid 0.36 2.26 0.36 2.26 0.36 2.26 sodium saltStabilizer 1.59 10.00 1.59 10.00 1.59 10.00 Water 4.53 28.47 4.53 28.474.53 28.47

TABLE 32 Co-Formulation of Ia-iii and imidacloprid M N O Amount AmountAmount Ingredient (g) Wt % (g) Wt % (g) Wt % MORWET D-425 0.36 2.26 0.362.26 0.36 2.26 Buffer 0.15 0.94 0.15 0.94 0.15 0.94 Propylene glycol0.90 5.66 0.90 5.66 0.90 5.66 ISOPAR M 0.36 2.26 0.36 2.26 0.36 2.26Antifoam 0.05 0.31 0.05 0.31 0.05 0.31 PLURONIC L-35 0.009 0.06 0.0090.06 0.009 0.06 Ia-iii 3.80 23.88 0.76 4.77 6.85 43.03 imidacloprid 3.8023.88 6.85 43.03 0.76 4.77 Humic acid 0.36 2.26 0.36 2.26 0.36 2.26sodium salt Stabilizer 1.59 10.00 1.59 10.00 1.59 10.00 Water 4.53 28.474.53 28.47 4.53 28.47

TABLE 33 Co-Formulation of Ia-i, imidacloprid, and metalaxyl PIngredient Amount (g) Wt % MORWET D-425 0.36 2.26% Propylene glycol 0.905.65% water 4.53 28.46% ISOPAR M 0.36 2.26% Humic Acid sodium salt 0.362.26% Ia-i 2.54 15.93% Imidacloprid 2.54 15.93% Metalaxyl 2.54 15.93%Antifoam 0.05 0.31% PLURONIC L-35 0.009 0.06% Buffer Solution 0.15 0.94%Stabilizer 1.59 9.99%

A small amount of each sample of co-formulation compositions A-P wastaken for particle size analysis. Particle size was measured by BeckmanCoulter particle size analyzer (Beckman Coulter LS 13 320 laserdiffraction particle size analyzer). The results are set forth below inTable 34.

TABLE 34 Particle Size for Co-Formulations Co- Mean Median Mean/MedianFormulation (μm) (μm) (μm) A 3.387 2.263 1.496 B 3.561 2.083 1.709 C3.146 2.213 1.422 D 3.757 2.525 1.488 E 5.697 3.442 1.655 F 2.993 2.0521.459 G 3.139 2.415 1.300 H 2.794 1.824 1.532 I 3.487 2.424 1.438 J3.514 2.142 1.640 K 5.046 2.473 2.040 L 3.569 2.380 1.499 M 5.491 4.0251.364 N 4.004 2.893 1.384 O 5.102 4.157 1.227 P 3.356 2.272 1.477

Example 27: Field Trial

In this Example, various nematicidal formulations were tested fornematicidal activity against soybean cyst nematode (SCN) in a soymicroplot field trial. The tested formulations included ACCELERON F/I, afungicide/insecticide seed treatment package available from MonsantoCompany and containing pyraclostrobin, metalaxyl, fluxapyroxad andimidacloprid, both alone and in combination with a composition inaccordance with the present invention containing the nematicidalcompound 3-phenyl-5-(thiophen-2-yl)-1,2,4-oxadiazole (Ia-i), inparticular Formulation 9 as described above in Table 27. Also tested wasthe combination of ACCLERON F/N, a fungicide+nematicide seed treatmentpackage available from Monsanto Company and containing pyraclostrobin,metalaxyl, fluxapyroxad, clothianidin and Bacillus firmus withFormulation 9.

Microplots containing a Stough fine sandy loam soil were fumigated withmethyl bromide. These plots were covered with a 114-p.m (4.5-mil) thickpolyethylene tarp for 72 hours. The tarp was removed and plots wereplanted 45 days later. Four seeds were planted in each microplot.Treatments were arranged in a randomized complete block design with fivereplications. Microplots were watered with drip irrigation as needed.Maximum and minimum weekly temperatures and amount of rainfall wererecorded for duration of the test.

A race 3 population of Heterodera glycines was increased on soybean(Coker 156) in a greenhouse. Light brown to tan colored cysts weredislodged from the roots with a strong water spray and collected onnested sieves with pore sizes of 850 and 250 I˜m. Cysts were placed into20-ml glass test tubes and crushed with a modified Seinhorst cystcrusher (21). The resultant suspension was passed through a 75-t×m-poresieve nested on a 28-˜m-pore sieve to remove broken cysts and debris.The inoculum was incorporated in the appropriate treatments by pipettingthe nematode suspension into 10 depressions 5 cm deep and 2 cm widewithin each microplot. Soil was then mixed with a garden hoe to a depthof 15 cm, obtaining an inoculum level of 2,000 eggs and J2/250 cm³ soil.

Plant stand, height and plant vigor evaluations were made at 20 to 40days after planting. The number of Heterodera glycines in each microplotwas determined at soybean maturity. Six soil cores 2.25-cm-d×15-cm deepwere collected from the soybean root zone in each microplot. Heteroderaglycines cysts were extracted from 250 cm a soil by sieving as describedfor inoculum production. J2 passing through the sieves used to collectcysts were extracted from the suspension using gravity-screening. Finalseparation of J2 in the fraction collected on the 28-um pore sieve wasby sucrose centrifugal flotation (sucrose specific gravity=1.13). Datawas reported as the number of J2s, cysts, eggs, J2+ eggs. Additionally,the treatment reproductive factor (Rf) was provided; Rf=final population(Pf)/the initial population (Pi).

Table 35 summarizes the SCN Reproductive Factor for the testedformulations.

TABLE 35 SCN Repro- Treat- ductive ment Factor Trt 1 ACCELERON F/Ipyraclostrobin, metalaxyl, 138 fluxapyroxad, imidacloprid Trt 2ACCELERON F/I + pyraclostrobin, metalaxyl, 73 Formulation 9 influxapyroxad, imidacloprid + Table 27 (0.25 Formulation 9 (0.25 mg/seed)mg/seed) Trt 3 ACCELERON F/I + pyraclostrobin, metalaxyl, 34 Formulation9 in fluxapyroxad, imidacloprid + Table 27 (0.50 Formulation 9 (0.50mg/seed) mg/seed) Trt 4 ACCELERON F/N + pyraclostrobin, metalaxyl, 28Formulation 9 in fluxapyroxad, clothianidin, Table 27 (0.50 Bacillusfirmus + mg/seed) Formulation 9 (0.50 mg/seed)

When introducing elements of the present invention or the preferredembodiments(s) thereof, the articles “a”, “an”, “the” and “said” areintended to mean that there are one or more of the elements. The terms“comprising”, “including” and “having” are intended to be inclusive andmean that there may be additional elements other than the listedelements.

In view of the above, it will be seen that the several objects of theinvention are achieved and other advantageous results attained.

As various changes could be made in the above products and methodswithout departing from the scope of the invention, it is intended thatall matter contained in the above description and the associateddrawings shall be interpreted as illustrative and not in a limitingsense.

What is claimed is:
 1. A method of preparing a nematicidal aqueoussuspension concentrate composition comprising an alkylaryl sulfonatedispersant, an organic solvent component comprising a paraffinichydrocarbon solvent, and a dispersed solid particulate phase comprisinga nematicidal component comprising a compound of Formula (I), Formula(II), or a salt thereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃, the method comprising: mixing thenematicidal component, the alkylaryl sulfonate dispersant, the organicsolvent component, and water to form an aqueous suspension; and wetmilling the aqueous suspension to produce a milled suspension having areduced particle size, wherein the median size of solid particulates inthe dispersed solid particulate phase of the milled suspension is lessthan about 10 μm.
 2. The method of claim 1 wherein the median size ofsolid particulates in the dispersed solid particulate phase of themilled suspension is less than about 5 μm.
 3. The method of claim 1wherein the median size of solid particulates in the dispersed solidparticulate phase of the milled suspension is from about 1 μm to about 5μm.
 4. The method of claim 1 wherein the mean particle size of solidparticulates in the dispersed solid particulate phase of the milledsuspension is less than about 20 μm.
 5. The method of claim 1 whereinthe mean particle size of solid particulates in the dispersed solidparticulate phase of the milled suspension is from about 0.5 μm to about10 μm.
 6. The method of claim 1 wherein the milled suspension has apolydispersity index of less than about
 10. 7. The method of claim 1wherein the milled suspension has a polydispersity index of from about 1to about
 2. 8. The method of claim 1 wherein the wet milling stepcomprises ball milling.
 9. The method of claim 1 wherein the nematicidalaqueous suspension concentrate composition further comprises a rheologymodifying agent.
 10. The method of claim 9 wherein the rheologymodifying agent comprises a humic substance.
 11. The method of claim 1wherein a secondary dispersant is mixed with the nematicidal component,the dispersant, and water prior to the wet milling step.
 12. The methodof claim 1 wherein an antifreeze agent is mixed with the nematicidalcomponent, the dispersant, and water prior to the wet milling step. 13.The method of claim 1 wherein a buffer solution is mixed with thenematicidal component, the dispersant, and water prior to the wetmilling step.
 14. The method of claim 13 wherein the pH of the aqueoussuspension during the wet milling step is less than about
 10. 15. Themethod of claim 1 wherein the milled suspension is combined with astabilizer component.
 16. The method of claim 1 wherein the milledsuspension is combined with a one or more biological control agents. 17.The method of claim 1 wherein the nematicidal aqueous suspensionconcentrate composition further comprises an antifoam agent.
 18. Themethod of claim 1 wherein the nematicidal component comprises a compoundof Formula (I) or a salt thereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃.
 19. The method of claim 18 whereinthe nematicidal component comprises a compound of Formula (Ia) or a saltthereof,

wherein R₁ and R₅ are independently selected from hydrogen, CH₃, F, Cl,Br, CF₃ and OCF₃; R₂ and R₄ are independently selected from hydrogen, F,Cl, Br, and CF₃; R₃ is selected from hydrogen, CH₃, CF₃, F, Cl, Br,OCF₃, OCH₃, CN, and C(H)O; R₇ and R₈ are independently selected fromhydrogen and F; R₉ is selected from hydrogen, F, Cl, CH₃, and OCF₃; andE is O or S.
 20. The method of claim 1 wherein the nematicidal componentcomprises a compound of Formula (II), or a salt thereof,

wherein A is selected from the group consisting of phenyl, pyridyl,pyrazyl, oxazolyl and isoxazolyl, each of which can be optionallyindependently substituted with one or more substituents selected fromthe group consisting of halogen, CF₃, CH₃, OCF₃, OCH₃, CN, and C(H)O;and C is selected from the group consisting of thienyl, furanyl,oxazolyl and isoxazolyl, each of which can be optionally independentlysubstituted with one or more substituents selected from the groupconsisting of F, Cl, CH₃, and OCF₃.